Conjugates derived from non-steroidal anti-inflammatory drugs and methods of use thereof in imaging

ABSTRACT

Conjugates derived from non-steroidal anti-inflammatory drugs (NSAIDs) and methods of use thereof are disclosed, useful for, inter alia, identifying and localizing the site of pathology and/or inflammation responsible for the sensation of pain in a patient; for identifying the sites of primary, secondary, benign, or malignant tumors; and for diagnosing infection or confirming or ruling out suspected infection. The NSAID-based conjugates contain an imaging moiety. The conjugates concentrate at sites of increased cyclooxygenase expression, thus revealing the sites of increased prostaglandin production, which is correlated with pain and inflammation, and correlated with tumor presence and/or location. Identifying areas of increased COX expressing can also aid in screening for infections.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims priority benefit of U.S. ProvisionalPatent Application No. 62/018,397, filed Jun. 27, 2014; of U.S.Provisional Patent Application No. 62/052,928, filed Sep. 19, 2014; andof U.S. Provisional Patent Application No. 62/109,544, filed Jan. 29,2015. The entire contents of those applications are hereby incorporatedby reference herein.

FIELD OF THE INVENTION

Conjugates derived from non-steroidal anti-inflammatory drugs (NSAIDs)and methods of use thereof are disclosed, useful for, inter alia,identifying and localizing the site of pathology and/or inflammationresponsible for the sensation of pain in a patient, sites of infection,and identifying and localizing the sites of tumor pathology, includingbenign, malignant, primary, and secondary tumors.

BACKGROUND OF THE INVENTION

It is important in medicine to identify the site of pathology in orderto properly screen for and/or treat a disease. Tumor screening for thepresence of tumors (e.g. for breast cancer, cervical cancer, coloncancer, prostate cancer, etc.) is very common. Some of the difficultieswith tumor screening are expense, patient's time, physician's time, andaccuracy. Also, many of the screening tests are not particularlyaccurate. For example, testing for prostate cancer using serum acidphosphatase or prostate specific antigen (PSA) is non-specific, andelevation of the marker in healthy individuals can be cause for anunnecessary surgery, a prostate biopsy. An additional example is MRIscreening for breast tumors, whose value has recently been questionedfor both insensitivity and occasional misinterpretation. In addition,the presence or absence of sentinel (metastatic) nodes is critical forthe optimal treatment of breast cancer. Low grade chondrosarcomas arenotoriously difficult to read by the pathologist, and frequently have tobe sent to multiple institutions for a diagnostic consensus. All ofthese examples suggest the need for improving detection for all benign,malignant, primary and secondary tumors. A rapid and non-invasive methodof localizing tumors would aid immensely in diagnosing and treating theunderlying cause. The growing tendency to understand tumors at themolecular level may also be guided by such improved non-invasivemethods.

Localization of pain is another area where identifying the site ofpathology is important for treatment; however, such localization isoften not straightforward. The unpleasant sensation of pain serves as anindicator of a disease or pathological state. Pain often occurs at thesite of pathology, and can be a helpful guide in determining diagnosisand appropriate treatment. However, in many cases, the area where apatient experiences pain may not be coincident with the area where theactual pathology has occurred. A classic example is sciatica, wherepressure on the sciatic nerve due to a herniated disc in the lower spinecan result in a sensation of pain in the leg, at a significant distancefrom the site of pathology. Another example is the difficulty indiagnosing pain in the chest or thorax, which can arise from multiplecauses, such as cardiac ischemia, gastroesophageal reflux, or pulmonaryembolism. In such cases, differential diagnosis requires a systematicprocess of elimination through tests and procedures until the causeand/or location of pathology is identified.

Screening for infectious diseases, particularly when a patient is stillasymptomatic, also poses difficulties. Medicaments and methods for suchscreening would prove useful in limiting outbreaks of diseases; earlytreatment of infected individuals; and avoiding unnecessary treatment orisolation for individuals who are suspected of being infected, but whoin actuality have not been infected, by a disease.

Because pathology is often accompanied by inflammation at the site ofthe pathology (which is not necessarily the site where pain isexperienced), rapid and non-invasive methods of localizing inflammationin a patient experiencing pain would aid immensely in diagnosing andtreating the underlying cause of the pain.

SUMMARY OF THE INVENTION

The current invention provides compounds and methods useful foridentification of areas of pathology, including tumors and inflammation,and screening for infections and sites of infections, via non-invasiveimaging. The compounds and methods can be used in both human andveterinary medicine.

In a first embodiment, the invention embraces conjugate comprising anon-steroidal anti-inflammatory drug (NSAID), a residue of a NSAID, or aderivative of a NSAID bonded or complexed to an imaging moiety whichcomprises a radioactive agent, wherein the radioactive agent is selectedfrom the group consisting of a gamma-ray emitter, an X-ray emitter, anda beta emitter; or a pharmaceutically acceptable salt thereof. In thisfirst embodiment, the imaging moiety can be optionally bonded to thenon-steroidal anti-inflammatory drug (NSAID), residue of a NSAID, orderivative of a NSAID via a linker. In a second embodiment, theradioactive agent in the first embodiment can be ^(99m)Tc, ⁵²Mn, ¹⁸⁶Reor ¹⁸⁸Re. In a third embodiment, the imaging moiety of the first orsecond embodiment can further comprise a chelating group which bonds orcomplexes to the radioactive agent; and the imaging moiety can beoptionally bonded to the non-steroidal anti-inflammatory drug (NSAID),residue of a NSAID, or derivative of a NSAID via a linker.

In a fourth embodiment, the imaging moiety comprising a chelating groupbonded or complexed to the radioactive agent of the third embodiment isof the form:

wherein M is selected from the group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Reor ¹⁸⁸Re

wherein each J is independently selected from the group consisting of NHand S,

R¹⁰¹, R¹⁰², and R¹⁰³ are independently selected from the groupconsisting of optionally substituted C₂-C₄ alkyl, and the NSAID, NSAIDresidue, or NSAID derivative is attached to the chelating group, eitherthrough a linker or directly, at

a) any J, R¹⁰¹, R¹⁰², or R¹⁰³ atom where a hydrogen atom can be replacedwith a bond to the linker (if present) or to the NSAID, NSAID residue,or NSAID derivative if no linker is present; or at

b) the nitrogen atom in the —R¹⁰¹—N—R¹⁰²— portion, forming a bondbetween that nitrogen and the linker (if present) or to the NSAID, NSAIDresidue, or NSAID derivative if no linker is present; or at

c) the nitrogen atom in the —R¹⁰²—N—R¹⁰³— portion, forming a bondbetween that nitrogen and the linker (if present) or to the NSAID, NSAIDresidue, or NSAID derivative if no linker is present;

or the imaging moiety comprising a chelating group bonded or complexedto the radioactive agent of the third embodiment is of the form:

wherein R_((ri)) is —CH₃ or —CH₂CH₃ and p is an integer between 0 and 4inclusive and the NSAID, NSAID residue, or NSAID derivative is attachedto the chelating group either through a linker or directly if no linkeris present, at any location on the cyclopentane ring which does not havea (R_(ri)) group.

In a fifth embodiment, in any of the first, second, third, or fourthembodiments, the NSAID, residue of an NSAID, or derivative of a NSAIDand the imaging moiety are bonded or complexed via a linker, where thelinker is selected from the group consisting of an optionallysubstituted C₁-C₁₀ hydrocarbylene group; an optionally substitutedC₂-C₁₀ heterohydrocarbylene group; and a linker of the form-L_(E)-R⁴-L_(F)-, where the linker -L_(E)-R⁴-L_(F)-can be in eitherorientation with respect to the NSAID, residue of an NSAID, orderivative of a NSAID and the imaging moiety (that is, either L_(E) orL_(F) can be attached to the NSAID, residue of an NSAID, or derivativeof a NSAID, and the other of L_(E) or L_(F) is attached to the imagingmoiety). In this fifth embodiment, L_(E) is absent or is selected fromthe group consisting of —NH—, —N(R⁸)—, and —C(═O)—, and R⁸ is optionallysubstituted C₁-C₄ alkyl, R⁴ is selected from the group consisting ofoptionally substituted C₁-C₁₀ hydrocarbylene, optionally substitutedC₂-C₁₀ heterohydrocarbylene, C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀ aryl-C₁-C₆ alkyl, and L_(F) isabsent or is a functional group selected from the group consisting of—(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—,—N(H)—(C═O)——N(R⁹)—(C═O)—(CH₂)—, —(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—,—N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—, —(CH═CH)—, or adivalent cycloalkyl or heterocyclic group, where R⁹ is selected from thegroup consisting of H and optionally substituted C₁-C₄ alkyl.

In a sixth embodiment, in any of the first, second, third, fourth, orfifth embodiments, the linker is selected from the group consisting of:—(NH)—(CH₂)_(n)—, —(NR_(a))—(CH₂)_(n)—, —(NH)—(CH₂)_(n)—(NH)—,—(NR_(a))—(CH₂)_(n)—(NR_(a))—, —(NH)—(CH₂CH₂)—(OCH₂CH₂)_(m)—(NH)—,—(NR_(a))—(CH₂CH₂)—(OCH₂CH₂)_(m)—(NR_(a))—,—(NH)—(CH₂CH₂)—((NH)CH₂CH₂)_(m)—(NH)—,—(NR_(a))—(CH₂CH₂)—((NH)CH₂CH₂)_(m)—(NR_(a))—, (—CH₂CH₂—O—)_(n),(—CH₂CH(CH₃)—O—)_(q), where R_(a) is (C₁-C₄ alkyl), n is an integer from1 to 10 inclusive, m is an integer from 1 to 4 inclusive, and q is aninteger from 1 to 3 inclusive,

—NH—(CH₂)₂—,—NH—(CH₂)₃—,—NH—(CH₂)₄—,—NH—(CH₂)₅—,—NH—(CH₂)₆—,—NH—(CH₂)₇—,—NH—(CH₂)₈—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—,—NH—(CH₂)₂—NH—,—NH—(CH₂)₃—NH—,—NH—(CH₂)₄—NH—,—NH—(CH₂)₅—NH—,—NH—(CH₂)₆—NH—,—NH—(CH₂)₇—NH—,—NH—(CH₂)₈—NH—,—NH—(CH₂)₃—C(CH₃)₂—NH—,—NH—(CH₂)₄—C(CH₃)₂—NH—,—NH—(CH₂)₅—C(CH₃)₂—NH—,—NH—(CH₂)₆—C(CH₃)₂—NH—,—NH—(CH₂)₇—C(CH₃)₂—NH—,—NH—(CH₂)₂—C(CH₃)₂—(CH₂)₂—NH—,—NH—(CH₂)₂—C(CH₃)₂—(CH₂)₂—,—NH—C(CH₃)₂—(CH₂)₃—,—NH—C(CH₃)₂—(CH₂)₄—,—NH—C(CH₃)₂—(CH₂)₅—,—NH—C(CH₃)₂—(CH₂)₆—,—NH—C(CH₃)₂—(CH₂)₇—,—NH—CH(CH₃)—(CH₂)₃—,—NH—CH(CH₃)—(CH₂)₄—,—NH—CH(CH₃)—(CH₂)₅—,—NH—CH(CH₃)—(CH₂)₆—,—NH—CH(CH₃)—(CH₂)₇—,—NH—CH(CF₃)—(CH₂)₃—,—NH—CH(CF₃)—(CH₂)₄—,—NH—CH(CF₃)—(CH₂)₅—,—NH—CH(CF₃)—(CH₂)₆—,—NH—CH(CF₃)—(CH₂)₇—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—,—NH—(CH₂)₂—NH—(C═O)—,—NH—(CH₂)₃—NH—(C═O)—,—NH—(CH₂)₄—NH—(C═O)—,—NH—(CH₂)₅—NH—(C═O)—,—NH—(CH₂)₆—NH—(C═O)—,—NH—(CH₂)₇—NH—(C═O)—,—NH—(CH₂)₈—NH—(C═O)—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—(C═O)—,—NH—(CH₂)₂—NH(C═O)—(CH₂)—,—NH—(CH₂)₃—NH(C═O)—(CH₂)—,—NH—(CH₂)₄—NH(C═O)—(CH₂)—,—NH—(CH₂)₅—NH(C═O)—(CH₂)—,—NH—(CH₂)₆—NH(C═O)—(CH₂)—,—NH—(CH₂)₇—NH(C═O)—(CH₂)—,—NH—(CH₂)₈—NH(C═O)—(CH₂)—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH(C═O)—(CH₂)—,—C(═O)—(CH₂)₂—,—C(═O)—(CH₂)₃—,—C(═O)—(CH₂)₄—,—C(═O)—(CH₂)₅—,—C(═O)—(CH₂)₆—,—C(═O)—(CH₂)₇—,—C(═O)—(CH₂)₈—,—C(═O)—(CH₂)₂—C(H)═C(H)—(CH₂)₂—,—C(═O)—(CH₂)₂—NH—,—C(═O)—(CH₂)₃—NH—,—C(═O)—(CH₂)₄—NH—,—C(═O)—(CH₂)₅—NH—,—C(═O)—(CH₂)₆—NH—,—C(═O)—(CH₂)₇—NH—,—C(═O)—(CH₂)₈—NH—,—C(═O)—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—,—NH—(CH₂)₂—O—(CH₂)₂—NH—,—NH—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—NH—,—NH—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—NH—,—NH—(CH₂)₂—NH—(CH₂)₂—NH—,—NH—(CH₂)₂—NH—(CH₂)₂—NH—(CH₂)₂—NH—,—NH—(CH₂)₂—NH—(CH₂)₂—NH—(CH₂)₂—NH—(CH₂)₂—NH—,—NH—(CH₂)₂—N(CH₃)—,—NH—(CH₂)₃—N(CH₃)—,—NH—(CH₂)₄—N(CH₃)—,—NH—(CH₂)₅—N(CH₃)—,—NH—(CH₂)₆—N(CH₃)—,—NH—(CH₂)₇—NCH₃)—,—NH—(CH₂)₈—NCH₃)—,—NH—(CH₂)₃—NH—(CH₂)₃—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—,—NH—CH₂—CF₂—(CH₂)₄—,—NH—CH₂—CF₂—(CH₂)₅——C(═O)—CF₂—(CH₂)₄—,—C(═O)—CF₂—(CH₂)₅—,

In a seventh embodiment, in any of the first, second, third, fourth,fifth, or sixth embodiments, the conjugate is of the formula:

or pharmaceutically acceptable salts thereof.

In this seventh embodiment, L_(E) is absent or is selected from thegroup consisting of —NH—, —N(R⁸)—, and —C(═O)—, and R⁸ is optionallysubstituted C₁-C₄ alkyl, R⁴ is selected from the group consisting ofoptionally substituted C₁-C₁₀ hydrocarbylene, optionally substitutedC₂-C₁₀ heterohydrocarbylene, C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀ aryl-C₁-C₆ alkyl, L_(F) isabsent or is a functional group selected from the group consisting of—(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—,—N(H)—(C═O)— —N(R⁹)—(C═O)—(CH₂)—, —(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—,—N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—, —(CH═CH)—, or adivalent cycloalkyl or heterocyclic group, where R⁹ is selected from thegroup consisting of H and optionally substituted C₁-C₄ alkyl; M isselected from the group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re or ¹⁸⁸Re;R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, and R¹⁷ are independently selectedfrom the group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkylsubstituted with fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl;or, independently of the other substituents, (R¹⁰ and R¹¹) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, (R¹² and R¹³) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, or (R¹⁴ and R¹⁵) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, or (R¹⁶ and R¹⁷) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, with the proviso that only oneof (R¹⁰ and R¹¹), (R¹² and R¹³), (R¹⁴ and R¹⁵), and (R¹⁶ and R¹⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring; R¹⁸, R¹⁹, R²⁰, R²¹, R²²,R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, and R²⁹ are independently selected fromthe group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substitutedwith fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or,independently of the other substituents, (R¹⁸ and R¹⁹) together with thecarbon to which they are attached form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, (R²⁰ and R²¹) together with the carbon to whichthey are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,or (R²² and R²³) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁴ and R²⁵) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁶ and R²⁷) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁸ and R²⁹) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,with the proviso that only one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²²and R²³), or (R²⁴ and R²⁵), or (R²⁶ and R²⁷), or (R²⁸ and R²⁹) togetherwith the carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring; or, independently of the othersubstituents, (R²² and R²³) together form an oxo group; Q is selectedfrom the group consisting of OH, NH₂, NH(C₁-C₄ alkyl), N(C₁-C₄alkyl)(C₁-C₄ alkyl), OCH₃, and OCH₂CH₃.

In an eighth embodiment, the conjugates of the seventh embodiment are ofthe form:

where R³⁸ is —O—(C₁-C₄) alkyl; R³⁹ is selected from the group consistingof H, F, Cl, Br, I, CH₃, and CF₃; R⁴⁰ is selected from the groupconsisting of H, CH₃, and CF₃; R⁴¹ and R⁴² are both H, or R⁴¹ is H andR⁴² is OH, or R⁴¹ and R⁴² together form an oxo group; R⁴³ is selectedfrom the group consisting of (C₁-C₁₀) hydrocarbylene and (C₂-C₁₀)heterohydrocarbylene containing one 0 atom, two 0 atoms, one (—S(═O₂))—group, two (—S(═O₂))— groups, or one 0 atom and one (—S(═O₂))— group,where the 0 atom or atoms of the heteroalkylene are not bonded directlyto a nitrogen atom, and where when more than one heteroatom is present,at least one carbon atom intervenes between the heteroatoms; M isselected from the group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re or ¹⁸⁸Re; orpharmaceutically acceptable salts thereof.

In a ninth embodiment, the conjugates of the eighth embodiment are ofthe formula:

where Re is ¹⁸⁶Re or ¹⁸⁸Re; or pharmaceutically acceptable saltsthereof.

In a terth embodiment, the conjugates of the eighth embodiment are ofthe formula:

wherein Tc is ^(99m)Tc; or pharmaceutically acceptable salts thereof.

In an eleventh embodiment, the conjugates of the eighth embodiment canbe selected from the group consisting of the compounds:

where Re is ¹⁸⁶Re or ¹⁸⁸Re; where Tc is ^(99m)Tc; and pharmaceuticallyacceptable salts thereof.

In a twelfth embodiment, the conjugates of the first or secondembodiment can be of the formula

(NSAID, NSAID residue, or NSAIDderivative)-(linker)-(chelator)-M-(terminal ligand)_(z1) or (NSAID,NSAID residue, or NSAID derivative)-(linker)-M-(terminal ligand)_(z2)where z1 is an integer between 0 and 4 inclusive; z2 is an integerbetween 0 and 5 inclusive; and -(linker)-(chelator)-M-(terminalligand)_(z1) or -(linker)-M-(terminal ligand)_(z2) is selected from thegroup consisting of:

where L_(E) is absent or is selected from the group consisting of —NH—,—N(R⁸)—, and —(C═O)—, and R⁸ is optionally substituted C₁-C₄ alkyl, withthe proviso that if the L_(E) moiety of the group -L_(F)-R⁴-L_(E)-wouldbe attached to a nitrogen atom, then L_(E) is absent; L_(G) is absent oris selected from the group consisting of —NH—, —N(R⁸)—, and —(C═O)—, andR⁸ is optionally substituted C₁-C₄ alkyl; R⁴ is selected from the groupconsisting of optionally substituted C₁-C₁₀ hydrocarbylene, optionallysubstituted C₂-C₁₀ heterohydrocarbylene, C₃-C₈ cycloalkyl, C₁-C₆alkyl-C₃-C₅ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ aryl-C₁-C₆ alkyl; R⁵ is selected from thegroup consisting of —OH, —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄alkyl), —OCH₃, and —OCH₂CH₃; R_(N) is H or (C₁-C₄ alkyl); L_(F) isabsent or is a functional group selected from the group consisting of—(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—,—N(H)—(C═O)——(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—,—N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl orheterocyclic group, where R⁹ is selected from the group consisting of Hand optionally substituted C₁-C₄ alkyl; and R¹⁸, R¹⁹, R²⁰, R²¹, R²²,R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, and R²⁹ are independently selected fromthe group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substitutedwith fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or,independently of the other substituents, (R¹⁸ and R¹⁹) together with thecarbon to which they are attached form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, (R²⁰ and R²¹) together with the carbon to whichthey are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,or (R²² and R²³) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁴ and R²⁵) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁶ and R²⁷) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁸ and R²⁹) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,with the proviso that only one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²²and R²³), or (R²⁴ and R²⁵), or (R²⁶ and R²⁷), or (R²⁸ and R²⁹) togetherwith the carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring; R³⁰, R³¹, R³², and R³³ areindependently selected from hydrogen and C₁-C₄ alkyl, or one pair of(R³⁰ and R³¹), (R³¹ and R³²), or (R³² and R³³), together with the atomsto which they are attached, form a six-membered aryl ring or afive-to-six membered heteroaryl ring; R³⁴, R³⁵, R³⁶, and R³⁷ areindependently selected from hydrogen and C₁-C₄ alkyl, or one pair of(R³⁴ and R³⁵), (R³⁵ and R³⁶), or (R³⁶ and R³⁷), together with the atomsto which they are attached, form a six-membered aryl ring or afive-to-six membered heteroaryl ring; M is selected from the groupconsisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re and ¹⁸⁸Re; R_((ri)) is —CH₃ or—CH₂CH₃ and p is an integer between 0 and 4 inclusive; and X is Cl orBr; or pharmaceutically acceptable salts thereof. In a furtherembodiment, R⁵ is selected from the group consisting of —NH₂, —NH(C₁-C₄alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —OCH₃, and —OCH₂CH₃; in yet afurther embodiment, R⁵ is selected from the group consisting of —NH₂,—OCH₃, and —OCH₂CH₃.

In a thirteenth embodiment, the -(linker)-(chelator)-M-(terminalligand)_(z1) or -(linker)-M-(terminal ligand)_(z2) group of theconjugates of the twelfth embodiment can be selected from the groupconsisting of:

where the substituents are as indicated in the twelfth embodiment, andpharmaceutically acceptable salts thereof.

In a fourteenth embodiment, the non-steroidal anti-inflammatory drug(NSAID), residue of a NSAID, or derivative of a NSAID in any of thefirst, second, third, fourth, fifth, sixth, seventh, eighth, ninth,terth, eleventh, twelfth, or thirteenth embodiments can be selected fromthe group consisting of:

where the atom marked with an asterisk * indicates an open valence atthat atom at which the NSAID, residue of the NSAID or derivative of theNSAID is attached to the remainder of the conjugate. In a fifteenthembodiment, the non-steroidal anti-inflammatory drug (NSAID), residue ofa NSAID, or derivative of a NSAID in any of the first, second, third,fourth, fifth, sixth, seventh, eighth, ninth, terth, eleventh, twelfth,or thirteenth embodiment can be selected from the group consisting of:

where the carbon atom or oxygen atom marked with an asterisk * indicatesan open valence at that atom at which the NSAID, residue of a NSAID, orderivative of a NSAID is attached to the remainder of the conjugate.

In a sixteenth embodiment, the conjugate of the first, second, third, orfourth embodiments can be selected from the group consisting of:

where Tc is ^(99m)Tc, Re is ¹⁸⁶Re or ¹⁸⁸Re; and pharmaceuticallyacceptable salts thereof.

In a seventeenth embodiment, the conjugate of any one of the first,second, or third embodiments can be selected from the group consistingof:

wherein Tc is ^(99m)Tc; and pharmaceutically acceptable salts thereof.

In an eighteenth embodiment, the conjugate of any one of the first,second, or third embodiments can be selected from the group consistingof:

wherein Tc is ^(99m)Tc; and pharmaceutically acceptable salts thereof.

In a nineteenth embodiment, the invention embraces a pharmaceuticalcomposition comprising one or more conjugates of any one of the first,second, third, fourth, fifth, sixth, seventh, eighth, ninth, terth,eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth,seventeenth, or eighteenth embodiments, and a pharmaceuticallyacceptable excipient.

In a twentieth embodiment, the invention embraces a method of imaging asite of pathology or suspected pathology in a subject, comprising:

a) administering one or more conjugates of any one of the first, second,third, fourth, fifth, sixth, seventh, eighth, ninth, terth, eleventh,twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, oreighteenth embodiments, or the pharmaceutical composition of thenineteenth embodiment, to the subject, where the radioactive agent ofthe conjugate comprises ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re or ¹⁸⁸Re; and

b) generating an image of the subject or an image of a portion of thesubject.

In a twentieth-first embodiment, the pathology or suspected pathology ofthe twentieth embodiment is a tumor or a suspected tumor.

In a twenty-second embodiment, the pathology or suspected pathology ofthe twentieth embodiment is pain (that is, the subject is suffering frompain).

In a twenty-third embodiment, the pathology or suspected pathology ofthe twentieth h embodiment is an infection or a suspected infection.

In a twenty-fourth embodiment, the invention embraces a conjugatedisclosed herein which has an IC₅₀ for inhibition of a cyclooxygenase ofless than about 2 micromolar, less than about 1 micromolar, preferablyless than about 0.5 micromolar, more preferably less than about 0.3micromolar, still more preferably less than about 0.1 micromolar, yetmore preferably less than about 50 nanomolar; in a twenty-fifthembodiment, the cyclooxygenase is COX-2. In a twenty-sixth embodiment,the invention embraces a conjugate of any one of the first, second,third, fourth, fifth, sixth, seventh, eighth, ninth, terth, eleventh,twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, oreighteenth embodiments which has an IC₅₀ for inhibition of acyclooxygenase of less than about 2 micromolar, less than about 1micromolar, preferably less than about 0.5 micromolar, more preferablyless than about 0.3 micromolar, still more preferably less than about0.1 micromolar, yet more preferably less than about 50 nanomolar; in atwenty-seventh embodiment, the cyclooxygenase is COX-2.

In some embodiments, the conjugates of the invention can be selectedfrom the group consisting of a conjugate illustrated in FIG. 1, FIG. 2,or FIG. 3; a rhenium-containing conjugate illustrated in FIG. 1, FIG. 2,or FIG. 3; a technetium-containing conjugate illustrated in FIG. 1, FIG.2, or FIG. 3; a technetium-99m-containing conjugate illustrated in FIG.1, FIG. 2, or FIG. 3; or a pharmaceutically acceptable salt thereof.

In further embodiments, the invention embraces a pharmaceuticalcomposition comprising one or more of the foregoing conjugates and apharmaceutically acceptable excipient.

In further embodiments, the invention embraces any of the conjugatesdisclosed herein, with the substitution of a non-radioactive agent forthe radioactive agent. Thus, for any of the generic structures orspecific conjugates disclosed herein containing ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re,or ¹⁸⁸Re, the invention also embraces those generic structures orspecific conjugates with a non-radioactive metal, such asnon-radioactive Re, such as ¹⁸⁵Re.

In further embodiments, the invention embraces any of the conjugatesdisclosed herein, with the removal of the radioactive agent. Thus, forany of the generic structures or specific conjugates disclosed hereincontaining ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re, the invention also embraces thosegeneric structures or specific conjugates without the metal, that is,with the uncomplexed or free chelator.

In further embodiments, the invention embraces the synthesis of any ofthe conjugates described herein, according to the protocols disclosedherein.

The invention also embraces methods of imaging a patient or subject. Inone embodiment, the invention embraces a method of imaging a site ofpathology or suspected pathology in a subject, comprising administeringone or more conjugates or compositions described herein, including anyof the foregoing embodiments, to the subject, wherein the radioactiveagent of the conjugate comprises ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re, or ¹⁸⁸Re; andgenerating an image of the subject or an image of a portion of thesubject. The pathology or suspected pathology in the subject can be atumor or a suspected tumor. The pathology or suspected pathology in thesubject can be an infection or a suspected infection. The subject can bea subject suffering from pain.

In further embodiments, the invention embraces any of the conjugates andcompositions as disclosed herein for use in imaging a subject or fordiagnosing a subject. In further embodiments, the invention embraces theuse in imaging or in diagnosis of any of the conjugates and compositionsas disclosed herein. In further embodiments, the invention embraces theuse of any of the conjugates and compositions as disclosed herein forthe manufacture of a medicament for use in imaging a subject, or for usein diagnostic purposes.

In further embodiments, the invention includes conjugates where theratio of (NSAID, NSAID residue, or NSAID derivative) to (chelatingmoiety) is 1:1.

Some embodiments described herein are recited as “comprising” or“comprises” with respect to their various elements. In alternativeembodiments, those elements can be recited with the transitional phrase“consisting essentially of” or “consists essentially of” as applied tothose elements. In further alternative embodiments, those elements canbe recited with the transitional phrase “consisting of” or “consists of”as applied to those elements. Thus, for example, if a composition ormethod is disclosed herein as comprising A and B, the alternativeembodiment for that composition or method of “consisting essentially ofA and B” and the alternative embodiment for that composition or methodof “consisting of A and B” are also considered to have been disclosedherein. Likewise, embodiments recited as “consisting essentially of” or“consisting of” with respect to their various elements can also berecited as “comprising” as applied to those elements. Finally,embodiments recited as “consisting essentially of” with respect to theirvarious elements can also be recited as “consisting of” as applied tothose elements, and embodiments recited as “consisting of” with respectto their various elements can also be recited as “consisting essentiallyof” as applied to those elements.

When a composition is described as “consisting essentially of” thelisted components, the composition contains the components expresslylisted, and may contain other components which do not substantiallyaffect the condition being treated. That is, the composition either doesnot contain any other components which do substantially affect thecondition being treated other than those components expressly listed;or, if the composition does contain extra components other than thoselisted which substantially affect the condition being treated, thecomposition does not contain a sufficient concentration or amount ofthose extra components to substantially affect the condition beingtreated. When a method is described as “consisting essentially of” thelisted steps, the method contains the steps listed, and may containother steps that do not substantially affect the condition beingtreated, but the method does not contain any other steps whichsubstantially affect the condition being treated other than those stepsexpressly listed.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates various ^(99m)Tc-containing conjugates useful in theinvention.

FIG. 2 illustrates various rhenium-containing conjugates.

FIG. 3 (3A-3S) illustrates various rhenium-containing and^(99m)Tc-containing conjugates.

DETAILED DESCRIPTION OF THE INVENTION

Identifying sites of pathology is important for proper diagnosis andtreatment of a patient. However, it can often be difficult to pinpointthe precise location of pathology. Extensive imaging and testing may berequired to accurately identify the source of pathology.

Tumor localization is an example of a condition where it can bedifficult to precisely identify the area of pathology, e.g., in apatient with metastatic adenocarcinoma who presents with clearmetastasis, but where the primary site of the malignancy is not known.The secondary sites of the tumor (metastases) are difficult to find inmany cancer cases. This problem also occurs with “benign tumors” such asgiant cell tumors, which rarely metastasize, and “quasi-malignanttumors” such as adamantinomas, which rarely metastasize early, but areknown to metastasize late in their course. Because the tumor locationcan be extremely difficult to find, a new test which could reveal alltypes of tumor cells would facilitate tumor searches, whether primarytumor sites or metastatic tumor sites, and help determine theappropriate treatment.

Pain is a common symptom in medicine, and is another condition where thesource of the pathology is not always readily apparent, despite thoroughphysical exams, laboratory studies, and radiologic studies and analysis.This is especially true for low back pain and abdominal pain. Pain inthe body results from various compounds produced and released at thesite of the injured area. These pain-producing compounds includebradykinins, prostaglandins, chemokines, histamine, and others.Importantly, the site at which the patient perceives the pain may not bethe site of the actual injury or pathology. The term “referred pain” isused to describe pain that is perceived by the patient at a sitedistinct from the pathology. Referred pain can complicate diagnosis,location of the actual site of pathology, and determination ofappropriate treatment.

Prostaglandins, especially the PG₂ group of prostaglandins, areover-expressed in tumor cells. Prostaglandins (such as the PG₂ group ofprostaglandins) are also strongly associated with the experience ofpain. Because prostaglandins are produced at the site of tumor location,actual injury, or pathology, identifying the site where prostaglandinsynthesis occurs will assist in locating the precise area of pathology.Biosynthesis of the PG₂ prostaglandins requires the cyclooxygenase (COX)enzyme. The cyclooxygenase enzyme exists in (at least) two isoforms,COX-1, which is expressed constitutively, but which may be upregulatedat sites of pain and inflammation, and COX-2, which is inducible byinflammatory stimuli. Both COX-1 and COX-2 are upregulated at tumorsites. Areas of high expression of cyclooxygenase will be associatedwith areas of high expression of prostaglandins, which in turn areassociated with the area of pathology. Thus, pinpointing areas of highcyclooxygenase expression will enable identification of the pathologicalarea.

The cyclooxygenase enzymes are readily inhibited by non-steroidalanti-inflammatory drugs (NSAIDs), which are sold over the counter inmost countries, and also often frequently prescribed by doctors. Thesenon-steroidal anti-inflammatory medicines include several pharmaceuticalclasses; each class has a number of specific drugs. If a NSAID drug isbonded or complexed to an imaging moiety, partial or total imaging ofthe patient provides a method of identifying sites of cyclooxygenaseoverexpression, prostaglandin synthesis, and inflammation, whichdetermines the site of pathology or injury. Thus, in one embodiment, theinvention encompasses conjugates of a NSAID with an imaging moiety,where said conjugate is suitable for imaging with an appropriate imagingmodality.

In addition to conjugates suitable for imaging, the invention alsoencompasses conjugates which are not used for imaging, but which areuseful surrogates for studying the chemical, biological, andpharmacokinetic properties of the conjugates suitable for imaging. Forexample, substitution of non-radioactive isotopes of rhenium (Re) for^(99m)Tc results in a conjugate which can be handled without the needfor radiation protection (the most abundant rhenium, isotope, ¹⁸⁷Re hasa half-life of on the order of 10¹⁰ years, and the second most abundantrhenium isotope, ¹⁸⁵Re is stable). Accordingly, preparation ofconjugates which have non-radioactive rhenium isotopes in place ofradioactive technetium isotopes can be useful for chemical, physical, invitro, and in vivo studies of compound properties in which the imagingproperties of the conjugate are not under study, such as studies oftoxicity and biological half-life, and the invention embraces both theconjugates suitable for imaging and their analogs which can be handledwithout radiation precautions.

The conjugates are also useful for diagnosis of infections. Infectionscause cells to overexpress the COX-1 and COX-2 enzymes. The patterndistribution of the cellular influence for the three major types ofinfections, bacterial, tuberculosis (TB), or viral, differ in majorways. Bacterial infections (not including TB) affect COX production inthe cells of most of the body's organs. The conjugates of the inventioncan be used for diagnosis of any bacterial infection, and areparticularly useful in abscess forming bacteria, in subjects or patientswith an organ-specific infection, and in aiding in diagnosis anddetermination of the cause of a fever of unknown origin (FUO). The organmost involved would produce more COX enzyme than the rest of the body'stissues, even though all tissues may show some increased activity.

TB infections can infect almost any organ, such as the lungs, thetestes, the spinal column (such as psoas abscess), etc. Scans conductedwith conjugates disclosed herein can help pinpoint the major locus of TBinfection, which is especially helpful in a subject or patient with apositive skin reaction to TB (such as a positive PPD test). The primarylocus for a TB infection would likely be at the site of the highestgamma count on a gamma camera when a gamma-emitting radioactive moietyis used in the conjugate.

Viral infections tend to first cause elevated COX production in thespleen to a great extent and in the stomach to a slightly lesser extent.The conjugates disclosed herein can thus be used for the screening ofasymptomatic patients infected with a virus. Patients are frequentlyinfectious even before they exhibit symptoms, such as patients withEbola virus and other viruses. An asymptomatic patient or subject whohas been exposed to such viruses, such Ebola virus, influenza viruses,or other viruses deemed sufficiently important for screening, or who hastraveled in areas where outbreaks of such viruses have occurred, can bescreened by administration of conjugates of the invention, followed byimaging. When a gamma-emitting radioactive moiety is used in theconjugate, a gamma scanner could detect signals above background (andthus increased COX expression) from at least the spleen and probably thestomach, indicating the presence of an infection.

Definitions

A “residue” of a non-steroidal anti-inflammatory drug (NSAID), referredto as an “NSAID residue” or “residue of a NSAID,” is a portion of theNSAID, where the portion of the NSAID retains its ability to bind tocyclooxygenase. Typically, a residue of a NSAID refers to the portion ofthe molecule left after removal of a hydrogen, a hydroxyl, a methyl, ora methoxy group from the NSAID. The residue is then bonded or complexedtogether with an imaging moiety. NSAID residues also include portions ofan NSAID that retains its ability to bind to cyclooxygenase, where theportion is further modified by the replacement of a hydrogen with ahalogen or a trifluoromethyl group, or by the replacement of a methylgroup with a trifluoromethyl group, or by the replacement of a hydroxylgroup with a methoxy group. In some embodiments, the residue can beconnected to a linker, which linker in turn is attached to a imagingmoiety, in order to bond or complex the NSAID residue with the imagingmoiety.

An NSAID derivative is an NSAID or NSAID residue which has been modifiedin one or more of the following manners:

a hydroxyl group has been replaced with a —O—(C₁-C₄ alkyl) group,preferably a methoxy group; a halogen; a C₁-C₄ alkyl group, preferably amethyl group; or a trifluoromethyl group;

a methyl group has been replaced with a trifluoromethyl group; a C₂-C₄alkyl group; a —O—(C₁-C₄ alkyl) group, preferably a methoxy group; ahalogen; or hydrogen;

a methoxy group has been replaced with a hydroxyl group, a —O—(C₂-C₄alkyl) group, a C₁-C₄ alkyl group, preferably a methyl group; a halogen,or a trifluoromethyl group;

a halogen has been replaced with a different halogen, a trifluoromethylgroup, a —O—(C₁-C₄ alkyl) group, preferably a methoxy group; a C₁-C₄alkyl group, preferably a methyl group; or hydrogen;

a hydrogen has been replaced with a halogen, a methyl group, or atrifluoromethyl group;

a —COOH group has been replaced with a —NH₂ group, or a —C(═O)—O— grouphas been replaced by a —NH— group;

a —COOH group has been replaced with a —CH₂NH₂ group, or a —C(═O)—O—group has been replaced by an —CH₂NH— group;

an amino group has been replaced by a carboxyl group;

a trifluoromethyl group has been replaced with a —CH₂NH₂ group or a—CH₂NH— group.

“Bonded or complexed” indicates that the NSAID, NSAID residue, or NSAIDderivative is associated with the imaging moiety in a manner such thatthe imaging moiety can be localized along with the NSAID, NSAID residue,or NSAID derivative to the intended site, such that the imaging moietycan indicate where the NSAID, NSAID residue, or NSAID derivative bondedor complexed to the imaging moiety has concentrated. “Bonded orcomplexed” embraces any manner of association sufficiently stable forthe foregoing purpose, whether such association is via a covalent bond,an ionic bond, a coordination bond (coordinate covalent bond or dativebond), a donor-acceptor complex, or any other association of NSAID,NSAID residue, or NSAID derivative and imaging moiety which has suitablestability. In one embodiment, “bonded or complexed” can indicate acovalent bond. In another embodiment, “bonded or complexed” can indicatea coordination bond. In another embodiment, “bonded or complexed” canindicate an ionic bond. In another embodiment, “bonded or complexed” canindicate a donor-acceptor complex.

“Alkyl” is intended to embrace a univalent saturated linear or branchedhydrocarbon chain having the number of carbon atoms specified, or if nonumber is specified, having 1 to 8 carbon atoms. “Alkylene” refers to asimilar group, which is divalent. “Optionally substituted” alkyl refersto either an unsubstituted alkyl group, or an alkyl group substitutedwith one or more substituents (such as one, two, three, four, or fivesubstituents) selected from the group consisting of —OH, —(C₁-C₄alkyl)-OH, halo, fluoro, chloro, bromo, iodo, —(C₁-C₄ alkyl), —(C₁-C₄)haloalkyl, —(C₁-C₄) perhaloalkyl, —O—(C₁-C₄ alkyl), —O—(C₁-C₄haloalkyl), —O—(C₁-C₄ perhaloalkyl), —(C₁-C₄) perfluoroalkyl,—(C═O)—(C₁-C₄) alkyl, —(C═O)—(C₁-C₄) haloalkyl, —(C═O)—(C₁-C₄)perhaloalkyl, —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl)(where each C₁-C₄ alkyl is chosen independently of the other), —NO₂,—CN, isocyano (NC—), oxo (═O), —C(═O)H, —C(═O)—(C₁-C₄ alkyl), —COOH,—C(═O)—O—(C₁-C₄ alkyl), —C(═O)NH₂, —C(═)ONH(C₁-C₄ alkyl), —C(═O)N(C₁-C₄alkyl)(C₁-C₄ alkyl) (where each C₁-C₄ alkyl is chosen independently ofthe other), —SH, —(C₁-C₄ alkyl)-SH, —S—(C₁-C₄ alkyl), —S(═O)—(C₁-C₄alkyl), —SO₂—(C₁-C₄ alkyl), and —SO₂—(C₁-C₄ perfluoroalkyl). Examples ofsuch substituents are —CH₃, —CH₂CH₃, —CF₃, —CH₂CF₃, —CF₂CF₃, —OCH₃,—NH(CH₃), —N(CH₃)₂, —SCH₃, and SO₂CH₃. “Optionally substituted alkylene”groups can be unsubstituted, or substituted in the same manner assubstituted alkyl groups.

“Cycloalkyl” is intended to embrace a univalent saturated cyclichydrocarbon chain having the number of carbon atoms specified, or if nonumber is specified, having 3 to 10 carbon atoms, or 3 to 8 carbonatoms, preferably 3 to 6 carbon atoms. “Cycloalkylene” refers to asimilar group, which is divalent. Cycloalkyl and cycloalkylene groupscan be unsubstituted, or substituted in the same manner as substitutedalkyl groups.

“Alkenyl” is intended to embrace a univalent linear or branchedhydrocarbon chain having at least one carbon-carbon double bond, andhaving the number of carbon atoms specified, or if no number isspecified, having 2 to 8 carbon atoms. “Alkenylene” refers to a similargroup, which is divalent. Alkenyl and alkenylene groups can beunsubstituted, or substituted in the same manner as substituted alkylgroups where chemically possible.

“Cycloalkenyl” is intended to embrace a univalent cyclic hydrocarbonchain having at least one carbon-carbon double bond and having thenumber of carbon atoms specified, or if no number is specified, having 4to 10 carbon atoms, or 4 to 8 carbon atoms, or 4 to 6 carbon atoms.“Cycloalkenylene” refers to a similar group, which is divalent.Cycloalkenyl and cycloalkenylene groups can be unsubstituted, orsubstituted in the same manner as substituted alkyl groups wherechemically possible.

“Alkynyl” is intended to embrace a univalent linear or branchedhydrocarbon chain having at least one carbon-carbon triple bond, andhaving the number of carbon atoms specified, or if no number isspecified, having 2 to 8 carbon atoms. “Alkynylene” refers to a similargroup, which is divalent. Alkynyl and alkynylene groups can beunsubstituted, or substituted in the same manner as substituted alkylgroups where chemically possible.

“Aryl” is defined as a univalent aromatic ring system. Aryl groupsinclude monocyclic aromatic rings and polycyclic aromatic ring systemscontaining the number of carbon atoms specified, or if no number isspecified, containing six to twenty carbon atoms. In other embodiments,aryl groups may contain six to ten carbon atoms. In some embodiments,aryl groups can be unsubstituted. In other embodiments, aryl groups canbe substituted with, for example, one, two, or three substituentsindependently selected from the group consisting of —OH, —(C₁-C₄alkyl)-OH, halo, fluoro, chloro, bromo, iodo, —(C₁-C₄ alkyl), —(C₁-C₄)haloalkyl, —(C₁-C₄) perhaloalkyl, —O—(C₁-C₄ alkyl), —O—(C₁-C₄haloalkyl), —O—(C₁-C₄ perhaloalkyl), —(C₁C₄) perfluoroalkyl,—(C═O)—(C₁-C₄) alkyl, —(C═O)—(C₁-C₄) haloalkyl, —(C═O)—(C₁-C₄)perhaloalkyl, —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl)(where each C₁-C₄ alkyl is chosen independently of the other), —NO₂,—CN, (NC—), —C(═O)H, —C(═O)—(C₁-C₄ alkyl), —COOH, —C(═O)—O—(C₁-C₄alkyl), —C(═O)NH₂, —C(═)ONH(C₁-C₄ alkyl), —C(═O)N(C₁-C₄ alkyl)(C₁-C₄alkyl) (where each C₁-C₄ alkyl is chosen independently of the other),—SH, —(C₁-C₄ alkyl)-SH and —S—C₁-C₄ alkyl. “Arylene” refers to a similargroup, which is divalent.

“Hydrocarbyl” is defined as a univalent hydrocarbon group, that is, agroup comprised of hydrogen and carbon, whether aliphatic or aromatic,acyclic or cyclic, or any combination of, or all of, aliphatic,aromatic, acyclic and cyclic. Hydrocarbyl groups have the number ofcarbon atoms specified, or if no number is specified, having 1 to 10carbon atoms. “Hydrocarbylene” refers to a similar group, which isdivalent. Hydrocarbyl and hydrocarbylene groups can be unsubstituted, orsubstituted in the same manner as substituted alkyl groups wherechemically possible.

“Heterocycle” or a “heterocyclic group” is defined as a ring systemwhich contains the number of carbon atoms specified, and one or moreheteroatoms (such as one to six heteroatoms, or one to threeheteroatoms, or one heteroatom), where heteroatoms include, but are notlimited to, oxygen, nitrogen, sulfur, and phosphorus. “Heteroaryl” isdefined as an aromatic ring system which contains the number of carbonatoms specified, and one or more heteroatoms (such as one to sixheteroatoms, or one to three heteroatoms, or one heteroatom), whereheteroatoms include, but are not limited to, oxygen, nitrogen, sulfur,and phosphorus; heteroaryl groups are a subset of heterocyclic groups.In some embodiments, heteroatoms for heterocyclyl and heteroaryl groupsare selected from the group consisting of oxygen and nitrogen. Invarious embodiments, heterocyclic groups may contain two to twentycarbon atoms and one to six heteroatoms, two to twelve carbon atoms andone to four heteroatoms, two to twelve carbon atoms and one to threeheteroatoms, two to ten carbon atoms and one to three heteroatoms, twoto six carbon atoms and one to three heteroatoms, or two to six carbonatoms and two to four heteroatoms. In some embodiments, heterocyclicgroups can be unsubstituted. In other embodiments, heterocyclic groupscan be substituted on any chemically possible valence with for example,one, two, or three substituents independently selected from the groupconsisting of —OH, —(C₁-C₄ alkyl)-OH, halo, fluoro, chloro, bromo, iodo,—(C₁-C₄ alkyl), —(C₁-C₄) haloalkyl, —(C₁-C₄) perhaloalkyl, —O—(C₁-C₄alkyl), —O—(C₁-C₄ haloalkyl), —O—(C₁-C₄ perhaloalkyl), —(C₁-C₄)perfluoroalkyl, —(C═O)—(C₁-C₄) alkyl, —(C═O)—(C₁-C₄) haloalkyl,—(C═O)—(C₁-C₄) perhaloalkyl, —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄alkyl)(C₁-C₄ alkyl) (where each C₁-C₄ alkyl is chosen independently ofthe other), —NO₂, —CN, (NC—), —C(═O)H, —C(═O)—(C₁-C₄ alkyl), —COOH,—C(═O)—O—(C₁-C₄ alkyl), —C(═O)NH₂, —C(═)ONH(C₁-C₄ alkyl), —C(═O)N(C₁-C₄alkyl)(C₁-C₄ alkyl) (where each C₁-C₄ alkyl is chosen independently ofthe other), —SH, —(C₁-C₄ alkyl)-SH and —S—C₁-C₄ alkyl. Examples ofheterocycles include aziridine, oxirane, oxetane, azetidine,pyrrolidine, pyrrole, tetrahydrofuran, furan, thiolane, thiophene,imidazolidine, imidazole, pyrazolidine, pyrazole, 1,2,3-triazole,1,2,4-triazole, piperidine, pyridine, pyran, piperazine, and morpholine.

A “heterohydrocarbyl” group is defined as a univalent hydrocarbyl group,where one or more of the carbon atoms have been independently replacedby a heteroatom at any chemically possible location, where heteroatomsinclude, but are not limited to, oxygen, nitrogen, sulfur, andphosphorus. Heterohydrocarbyl groups have the number of carbon atomsspecified, or if no number is specified, having 1 to 10 carbon atoms,and also at least one heteroatom, such as 1 to 5 heteroatoms, 1 to 4heteroatoms, 1 to 3 heteroatoms, 1 to 2 heteroatoms, or one heteroatom.“Heterohydrocarbylene” refers to a similar group, which is divalent.Heterohydrocarbyl and heterohydrocarbylene groups can be unsubstituted,or substituted in the same manner as substituted alkyl groups wherechemically possible. Examples of heterohydrocarbyl andheterohydrocarbylene groups include, but are not limited to, ethyleneglycol and polyethylene glycol moieties, such as (—CH₂CH₂—O)n—H (amonovalent heterohydrocarbyl group) and (—CH₂CH₂—O—)_(n) (a divalentheterohydrocarbylene group) where n is an integer from 1 to 12inclusive, and propylene glycol and polypropylene glycol moieties, suchas (—CH₂CH(CH₃)—O—)_(n)—H (a monovalent heterohydrocarbyl group) and(—CH₂CH(CH₃)—O—)_(n)— (a divalent heterohydrocarbylene group) where n isan integer from 1 to 12 inclusive. For heterohydrocarbyl groups, whenmore than one heteroatom is present, at least one carbon atom intervenesbetween any two heteroatoms.

The various groups described above can be attached to the remainder ofthe molecule at any chemically possible location on the fragment,including attachment via a substituent when the group is substituted.For the purposes of drawing the structures, groups are typicallyattached by replacement of a hydrogen, hydroxyl, methyl, or methoxygroup on a “complete” molecule to generate the appropriate fragment, anda bond is drawn from the open valence on the fragment to the remainderof the molecule. For example, attachment of the heteroalkyl group—CH₂—O—CH₃ proceeds by removal of a hydrogen from one of the methylgroups of CH₃—O—CH₃, to generate the heteroalkyl fragment —CH₂—O—CH₃,from which a bond is drawn from the open valence to the remainder of themolecule.

Reference to “about” a value or parameter herein includes (anddescribes) variations that are directed to that value or parameter perse. For example, description referring to “about X” includes descriptionof “X”.

The terms “a” or “an,” as used in herein means one or more, unless thecontext clearly indicates otherwise.

By “subject,” “individual,” or “patient” is meant an individualorganism, preferably a vertebrate, more preferably a mammal, mostpreferably a human.

The description is intended to embrace all salts of the compoundsdescribed herein, as well as methods of using such salts of thecompounds. In one embodiment, the salts of the compounds comprisepharmaceutically acceptable salts. Pharmaceutically acceptable salts arethose salts which can be administered as drugs or pharmaceuticals tohumans and/or animals and which, upon administration, retain at leastsome of the biological activity of the free compound (neutral compoundor non-salt compound). The desired salt of a basic compound may beprepared by methods known to those of skill in the art by treating thecompound with an acid. Examples of inorganic acids include, but are notlimited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitricacid, and phosphoric acid. Examples of organic acids include, but arenot limited to, formic acid, acetic acid, propionic acid, glycolic acid,pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid,fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid,mandelic acid, sulfonic acids, and salicylic acid. Salts of basiccompounds with amino acids, such as aspartate salts and glutamate salts,can also be prepared. The desired salt of an acidic compound can beprepared by methods known to those of skill in the art by treating thecompound with a base. Examples of inorganic salts of acid compoundsinclude, but are not limited to, alkali metal and alkaline earth salts,such as sodium salts, potassium salts, magnesium salts, and calciumsalts; ammonium salts; and aluminum salts. Examples of organic salts ofacid compounds include, but are not limited to, procaine, dibenzylamine,N-ethylpiperidine, N,N′-dibenzylethylenediamine, and triethylaminesalts. Salts of acidic compounds with amino acids, such as lysine salts,can also be prepared. For lists of pharmaceutically acceptable salts,see, for example, P. H. Stahl and C. G. Wermuth (eds.) “Handbook ofPharmaceutical Salts, Properties, Selection and Use” Wiley-VCH, 2011(ISBN: 978-3-90639-051-2). Several pharmaceutically acceptable salts arealso disclosed in Berge, J. Pharm. Sci. 66:1 (1977).

The invention also includes, where chemically possible, allstereoisomers and geometric isomers of the compounds, includingdiastereomers, enantiomers, and cis/trans (E/Z) isomers. The inventionalso includes mixtures of stereoisomers and/or geometric isomers in anyratio, including, but not limited to, racemic mixtures. Unlessstereochemistry is explicitly indicated in a structure, the structure isintended to embrace all possible stereoisomers of the compound depicted.If stereochemistry is explicitly indicated for one portion or portionsof a molecule, but not for another portion or portions of a molecule,the structure is intended to embrace all possible stereoisomers for theportion or portions where stereochemistry is not explicitly indicated.

Unless a specific isotope is indicated, the invention includes allisotopologues of the compounds disclosed herein, such as, for example,deuterated derivatives of the compounds (where H can be ²H, i.e., D).

The following abbreviations may be used herein:

-   ˜ about-   +ve or pos. ion positive ion-   Δ heat-   AA Arachidonic acid-   Ac Acetyl-   ACN acetonitrile-   Ac₂O acetic anhydride-   aq aqueous-   AcOH acetic acid-   Bn benzyl-   Boc tert-butyloxycarbonyl-   BOP-Cl Bis(2-oxo-3-oxazolidinyl)phosphinic chloride-   BSA bovine serum albumin-   Bu butyl-   Bz benzoyl-   Calcd or Calc'd calculated-   Conc. concentrated-   COX cyclooxygenase, prostaglandin-endoperoxide synthase-   d day(s)-   DCE dichloroethane-   DCM dichloromethane-   DEA diethylamine-   DEAD Diethyl azodicarboxylate-   DIEA or DIPEA diisopropylethylamine-   DMAP 4-dimethylaminopyridine-   DME 1,2-dimethoxyethane-   DMF N,N-dimethylformamide-   DMSO dimethyl sulfoxide-   EDC or EDCI N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide-   EIA Enzyme immunoassay-   eq equivalent-   ESI or ES electrospray ionization-   Et ethyl-   Et₂O diethyl ether-   Et₃N triethylamine-   EtOAc ethyl acetate-   EtOH ethyl alcohol-   g gram(s)-   h hour(s)-   HATU 0-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   HBTU    0-benzotriazole-N,N,N′,N′-tetramethyl-uronium-hexafluorophosphate-   Hex hexanes-   HMPA hexamethylphosphoramide-   HOAt 1-hydroxy-7-azabenzotriazole-   HOBt hydroxybenzotriazole-   HPLC high pressure liquid chromatography-   IPA or iPrOH isopropyl alcohol-   KOAc potassium acetate-   LCMS, LC-MS or LC/MS liquid chromatography mass spectrometry-   LDA lithium diisopropylamide-   LHMDS or LiHMDS lithium hexamethyldisilazide-   M molar (mol L⁻¹)-   Me methyl-   MeCN acetonitrile-   Mel iodomethane-   MeOH methyl alcohol-   mg milligram(s)-   min minute(s)-   mL milliliter(s)-   M mole(s)-   MS mass spectrometry-   MsC1 methanesulfonyl chloride-   MTBE or MtBE methyl tert-butyl ether-   m/z mass-to-charge ratio-   NaHMDS sodium hexamethyldisilazide-   NaOtBu sodium tert-butoxide-   NB S N-bromosuccinimide-   NCS N-chlorosuccinimide-   nBuLi n-butyl lithium-   NMO N-methylmorpholine-N-oxide-   NMP 1-methyl-2-pyrrolidinone-   NMR nuclear magnetic resonance-   PE Petroleum ether, CAS Number: 101316-46-5-   PG Prostaglandin or prostaglandins-   PGE2 Prostaglandin E2-   PGF2α Prostaglandin F2α-   PGH2 Prostaglandin H2-   PBS phosphate buffered saline-   PMB para-methoxybenzyl, 4-methoxybenzyl-   Pr propyl-   Prep-HPLC Preparative high pressure liquid chromatography-   ppm parts per million-   p-tol para-toluoyl-   rac racemic-   RP-HPLC or RPHPLC reversed phase high pressure liquid chromatography-   RT or rt or r.t. room temperature-   sat. or sat′d or satd saturated-   TBAF tetrabutylammonium fluoride-   TBDMS tert-butyldimethylsilyl-   TBDMS-Cl tert-butyldimethylsilyl chloride-   TBDPS tert-butyldiphenylsilyl-   TEMPO (2,2,6,6-tetramethylpiperidin-1-yl)oxidanyl-   tert or t tertiary-   TFA triflouroacetic acid-   THF tetrahydrofuran-   TLC thin layer chromatography-   TMS trimethylsilyl or trimethylsilane-   t_(R) retention time-   tBuOH tert-butyl alcohol-   v/v volume per volume

Non-Steroidal Anti-Inflammatory Drugs (NSAIDs)

Non-steroidal anti-inflammatory drugs (NSAIDs) are commonly used totreat pain and inflammation, which occur due to many diseases, such asthose involving tumors. NSAIDs which can be used in the inventioninclude:

salicylates, such as aspirin (acetylsalicylic acid), diflunisal,salsalate, and choline magnesium trisalicylate;

propionic acid derivatives, such as ibuprofen, dexibuprofen, naproxen,fenoprofen, ketoprofen, dexketoprofen, flurbiprofen, oxaprozin, andloxoprofen;

acetic acid derivatives, such as indomethacin, tolmetin, sulindac,etodolac, ketorolac, diclofenac, aceclofenac, and nabumetone;

enolic acid (oxicam) derivatives, such as piroxicam, meloxicam,tenoxicam, droxicam, lornoxicam, and isoxicam;

anthranilic acid derivatives (fenamates), such as mefenamic acid,meclofenamic acid, flufenamic acid, and tolfenamic acid;

selective COX-2 inhibitors (coxibs, several of which are not approved,or have been withdrawn, in various countries), such as celecoxib,rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, and firocoxib(firocoxib approved for equine and canine use only);

sulfonanilides such as nimesulide (which should be used cautiously dueto the risk of liver damage); and

licofelone, which is both a cyclooxygenase and lipoxygenase inhibitor.

When forming a conjugate of a NSAID compound, typically a bond to anatom or atoms on the NSAID compound is replaced with a bond to a linkinggroup in order to facilitate bonding or complexing the NSAID, NSAIDresidue, or NSAID derivative to the imaging moiety. Thus, for example,linking an amine-containing linker to a carboxylic acid-containing NSAIDcompound can be accomplished by condensing the amine and carboxylic acidto form an amide group. The bond to the —OH portion of the —COOH grouphas been replaced with a bond to the nitrogen atom of theamine-containing linker. “NSAID residue” refers to the portion of theNSAID molecule that remains and which is attached to the linker, and, asdefined previously, retains its ability to bind to cyclooxygenase. TheNSAID residue need not bind to COX with the same affinity or specificityas the NSAID from which the residue is derived, as long as sufficientbinding affinity remains for the purposes of the invention. A derivativeof a NSAID includes an NSAID or NSAID residue that has been modified asdefined herein, and the NSAID derivative portion of the conjugate neednot bind to COX with the same affinity or specificity as the NSAID whichwas modified to form the derivative, as long as sufficient bindingaffinity remains for the purposes of the invention.

Linker groups and linker/chelator groups for attachment of imagingmoieties to NSAID compounds; general and exemplary forms of conjugates

An imaging moiety can be bonded or complexed to a NSAID compound byusing an appropriate linker (also referred to as a linking group). Thelinker must be capable of being bonded or complexed to both the NSAIDcompound, or to a residue or derivative of the NSAID compound, andbonded or complexed to the imaging moiety, for the purpose of localizingthe imaging moiety to the site where the NSAID compound or residue orderivative of the NSAID compound concentrates when administered to apatient. In various non-limiting embodiments, a linker group covalentlybound to both the NSAID and the imaging moiety, or a linker groupcovalently bound to the NSAID and bonded or complexed via a coordinationbond to the imaging moiety, or a linker group covalently bound to theNSAID and bonded or complexed via an ionic bond to the imaging moiety,can be used. The linker should preferably be bonded or complexed to theNSAID molecule or residue or derivative of the NSAID at a site asdistant as possible from the region of the NSAID or residue orderivative of the NSAID that binds to cyclooxygenase, in order topreserve the ability of the NSAID or residue or derivative of the NSAIDto bind to cyclooxygenase. However, any attachment to the NSAID moleculeor residue or derivative of the NSAID, which attachment does not renderthe conjugate unsuitable for its purpose, can be used.

The linker also needs to bind or complex with the imaging moiety. Oneparticularly useful imaging moiety is the metastable isotope oftechnetium-99, referred to as ^(99m)Tc, Tc99m, or Tc-99m. ^(99m)Tcdecays to ⁹⁹Tc with a half-life of about 6 hours, via gamma emission atan energy of about 140 keV. ^(99m)Tc is often used in a complex with sixmethoxyisobutylisonitrile (MIBI) ligands, which is often referred to as^(99m)Tc-MIBI, technetium Tc99m Sestamibi, or ^(99m)Tc sestamibi (soldas CARDIOLITE, a registered trademark of Lantheus Medical Imaging, Inc.,North Billerica, Mass.). This compound has the following structure:

Modification of a NSAID with a linker such as

enables the linker to bind to a radioactive tracer moiety, such as^(99m)Tc, via the isocyano group, while the remaining open valence onthe linker will allow attachment of a NSAID or a residue or derivativeof a NSAID. For example, the structure below can be attached to aradioactive tracer moiety, such as ^(99m)Tc:

where R¹, R², and R³ can be independently selected from the groupconsisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted withhydroxy, fluoro, C₁-C₄ alkyl, or C₃-C₆ cycloalkyl, or R¹ and R² togetherwith the carbon to which they are attached form a C₃-C₈ cycloalkyl ringor heterocycloalkyl ring, while n is an integer selected from 0 to 4,inclusive.

The linker itself may be capable of bonding to or complexing theradioactive agent, as with the methoxyisobutylisonitrile ligands abovewhich directly bond to ^(99m)Tc. In other embodiments, the imagingmoiety comprises a chelating group and the radioactive agent, and mayfurther comprise additional groups, such as ligands, bound or complexedto the radioactive agent. Chelating groups can comprise a number ofligand systems known to the skilled artisan, such as those described ine.g. Jürgens, S. et al., J. Organomet. Chem. (2014), 751, 83-89 and Liu,G. et al. Anticancer Agents Med. Chem. (2007), 7(3): 367-377.

In some embodiments, when a linker is used to link a NSAID, NSAIDresidue, or NSAID derivative to an imaging moiety, the linker can beselected from the group consisting of an optionally substituted C₁-C₄₀hydrocarbylene group; an optionally substituted C₂-C₄₀heterohydrocarbylene group; and a linker of the form -L_(E)-R₄-L_(F)-,where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, R⁸ is optionally substituted C₁-C₄ alkyl, R⁴ is selectedfrom the group consisting of optionally substituted C₁-C₄₀hydrocarbylene and optionally substituted C₂-C₄₀ heterohydrocarbylene,C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₅ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀aryl-C₁-C₆ alkyl; and L_(F) is absent or is a functional group selectedfrom the group consisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—,—N(R⁹)—(C═O)—, —(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—,—N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl orheterocyclic group such as 1,2,3-triazole or 1,2,4-triazole, where R⁹ isselected from the group consisting of H and optionally substituted C₁-C₄alkyl.

In other embodiments, when a linker is used to link a NSAID, NSAIDresidue, or NSAID derivative to an imaging moiety, the linker can beselected from the group consisting of an optionally substituted C₁-C₁₀hydrocarbylene group; an optionally substituted C₂-C₁₀heterohydrocarbylene group; and a linker of the form -L_(E)-R⁴-L_(F)-;where L_(E) is absent or is selected from the group consisting of —NH—,—N(R⁸)—, and —C(═O)—, and R⁸ is optionally substituted C₁-C₄ alkyl, R⁴is selected from the group consisting of optionally substituted C₁-C₁₀hydrocarbylene, optionally substituted C₂-C₁₀ heterohydrocarbylene,C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀aryl-C₁-C₆ alkyl, and L_(F) is absent or is a functional group selectedfrom the group consisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—,—N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—, —N(R⁹)—(C═O)—(CH₂)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O)N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl.

A useful chelating group for preparation of conjugates is thetetradentate mertiatide group(N—[N—[N-[(acetylthio)acetyl]glycyl]glycyl]-glycine; Fritzberg et al.,J. Nuclear Medicine (1986) 27(1):111-116), having the structure:

For example, the following structure shows metal complexed with a groupderived from mertiatide to form an imaging moiety:

where M indicates a cationic metal ion, where the metal ion also bearsan oxo group (for example, M=0 can be ^(99m)Tc=O or Re═O). Thecarboxylic acid functionality of the complex allows for a wide range ofcoupling reactions in order to attach the imaging moiety to theremainder of the molecule. Typically, the mertiatide peptide is coupledvia its carboxylate group prior to introduction of the metal ion ormetal-oxo ion. Once the mertiatide peptide is complexed to the metal ion(or metal-oxo ion), it can be referred to either as “mertiatidecomplexed to the metal ion (or metal-oxo ion),” or as a group derivedfrom mertiatide. That is, a group bound or complexed to a metal ion ormetal-oxo ion which is referred to as “derived” from a specific moietyrefers to the group after it has complexed with the metal ion ormetal-oxo ion. Alternatively, the group which binds or complexes to themetal ion or metal-oxo ion can be referred to by the same name in itsfree, uncomplexed form and in its complexed form with the metal; forexample, “mertiatide group” can refer to both a mertiatide group withouta complexed metal or metal-oxo ion, and also a mertiatide group whichhas complexed with a metal or metal-oxo ion.

A mertiatide-based group of the form:

such as

can be used in conjugates of the invention, where L_(E) is absent or isselected from the group consisting of —NH— and —N(R⁸)—, where R⁸ isoptionally substituted C₁-C₄ alkyl, R⁴ is selected from the groupconsisting of optionally substituted C₁-C₃₀ hydrocarbylene, optionallysubstituted C₂-C₃₀ heterohydrocarbylene group (such as C₁-C₁₂ alkylene),C₃-C₅ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀aryl-C₁-C₆ alkyl; and L_(F) is absent or is a functional group selectedfrom the group consisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—,—N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—, —(SO₂) N(R⁹)—, —N(R⁹)—(SO₂)—,—N(R⁹) (C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, and —O—(C═O)N(R⁹)— where onevalence of L_(F) (when L_(F) is present) is attached to the R⁴ group andthe other valence is attached to the NSAID or residue or derivative of aNSAID; and where each R⁹ is independently selected from the groupconsisting of H and optionally substituted C₁-C₄ alkyl. In oneembodiment, R⁹ is H. The substituents R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶,and R¹⁷ are independently selected from the group consisting ofhydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted with fluoro, hydroxy,—O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or, independently of the othersubstituents, (R¹⁰ and R¹¹) together with the carbon to which they areattached independently form a C₃-C₈ cycloalkyl ring or heterocycloalkylring, (R¹² and R¹³) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R¹⁴ and R¹⁵) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R¹⁶ and R¹⁷) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring. Insome embodiments, only one of (R¹⁰ and R¹¹), (R¹² and R¹³), (R¹⁴ andR¹⁵), and (R¹⁶ and R¹⁷) together with the carbon to which they areattached independently form a C₃-C₈ cycloalkyl ring or heterocycloalkylring; that is, only one of the four pairs of substituents forms a spiroring, and the remaining three pairs do not form a spiro ring.

TechneScan MAG3™ is a commercially available kit for preparing^(99m)Tc-mertiatide conjugates, and is sold by MallinckrodtPharmaceuticals (catalog number N096B0, National Drug Code 00019N096B0),675 McDonnell Blvd., St. Louis, Mo. 63042, USA. The kit vials containbetiatide, stannous chloride dihydrate (SnCl₂.2H₂O), sodium tartratedihydrate, and lactose monohydrate. Betiatide, which isN-[(benzoylthio)acetyl]glycylglycylglycine, reacts with sodiumpertechnetate ^(99m)Tc to form ^(99m)Tc-mertiatide.

Yet another useful chelating group for preparation of conjugates isbased on the tetradentate ethylenedicysteine group:

or, in stereospecific form using L-cysteine (having the R configurationat the alpha carbon):

The following structure shows a metal ion complexed with a linkerderived from ethylenedicysteine:

where M indicates a cationic metal ion, where the metal ion also bearsan oxo group (for example, M=0 can be ^(99m)Tc═O or Re═O). Again, thecarboxylic acid functionalities of the complex allows for a wide rangeof coupling reactions. One of the carboxylic acid functionalities can beblocked, for example as an amide:

while the remaining carboxylic acid group can be used for attachment tothe remainder of the molecule.

An ethylenedicysteine-based linker-imaging moiety of the form:

can be used in conjugates of the invention, where L_(E) is absent or isselected from the group consisting of —NH— and —N(R⁸)—, where R⁸ isoptionally substituted C₁-C₄ alkyl, R⁴ is selected from the groupconsisting of optionally substituted C₁-C₃₀ hydrocarbylene, optionallysubstituted C₂-C₃₀ heterohydrocarbylene group (such as C₁-C₁₂ alkylene),C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl C₁₋₆ aryl-C₁-C₆ alkyl, and C₁-C₆alkyl-C₆-C₁₀ alkyl, R⁵ is —OH, —OR⁶, —NH₂, —NHR⁶, or —NR⁶R⁷, R⁶ and R⁷are independently selected from C₁-C₄ alkyl, and L_(F) is absent or is afunctional group selected from the group consisting of —(C═O)—, —O—,—N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—, —(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)—(C═O)—N(R⁹)—, —N(R⁹)—(C═O)—O—, and—O—(C═O)N(R⁹)— where one valence of L_(F) (when L_(F) is present) isattached to the R⁴ group and the other valence is attached to the NSAIDor residue or derivative of a NSAID; and where each R⁹ is independentlyselected from the group consisting of H and optionally substituted C₁-C₄alkyl. In one embodiment, R⁹ is H. The substituents R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, and R²⁵ are independently selected from the groupconsisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted withfluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or, independentlyof the other substituents, (R¹⁸ and R¹⁹) together with the carbon towhich they are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkylring, (R²⁰ and R²¹) together with the carbon to which they are attachedform a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²² and R²³)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁴ and R²⁵)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring. In some embodiments,only one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²² and R²³), or (R²⁴ andR²⁵) together with the carbon to which they are attached independentlyform a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring; that is, only oneof the four pairs of substituents forms a spiro ring, and the remainingthree pairs do not form a spiro ring. In a further embodiment, R⁵ isselected from the group consisting of —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄alkyl)(C₁-C₄ alkyl), —OCH₃, and —OCH₂CH₃; in yet a further embodiment,R⁵ is selected from the group consisting of —NH₂, —OCH₃, and —OCH₂CH₃.

It should be noted that the mertiatide fragment:

is a C₁₀ heterohydrocarbyl group (specifically, C₁₀ heteroalkyl) withthree nitrogen atoms and a sulfur atom in the chain, and five oxosubstituents, while the ethylenedicysteine fragment:

is a C₆ heterohydrocarbyl group (specifically, C₆ heteroalkyl);following the chain from the first thiol-substituted carbon to thesecond thiol-substituted carbon, the ethylenedicysteine fragment has twonitrogen atoms in the chain, two —SH substituents on the first and lastcarbons in the chain, one —COOH substituent, and a second —COOHsubstituent which is linked to the remainder of the molecule by removalof the —OH group of the carboxyl. Alternatively, the ethylenedicysteinefragment can be considered as a 3,6-diazaoctane (i.e., heteroalkyl) withthiol substituents on the 1 and 8 carbons, a carboxy substituent on the7 carbon, and a —C(═O)—OH or —C(═O)—H moiety on the 2 carbon, where the—OH or —H, respectively, has been removed for attachment to theremainder of the molecule.

In one embodiment, where the linker(s) is (are) capable of directlybinding to or chelating the metal of the imaging moiety, the inventioncomprises compounds of the formula:

[(NSAID, NSAID residue, or NSAID derivative)-(linker)]_(x)-M-(terminalligand)_(z1)

where “NSAID, NSAID residue, or NSAID derivative” refers to a NSAID or aresidue or derivative of a NSAID, M is selected from the groupconsisting of ^(99m)Tc, ⁵²Mn, and Re, x is an integer between 1 and 6inclusive, z1 is an integer between 0 and 5 inclusive, and the sum of xand z1 is less than or equal to 6, and “terminal ligand” is a ligandthat coordinates to M, but which does not have a NSAID attached to it.The linker or linkers may be monodentate or polydentate for M. Theterminal ligand or ligands may be monodentate or polydentate for M. Whenmore than one terminal ligand is present, all of the ligands can beidentical groups, all of the ligands can be different groups, or some ofthe ligands can be identical groups and some can be different groups. Itshould be noted that some monodentate ligands, such as the oxo ligand,═O, can occupy more than one valence on the metal ion; that is, therecan be multiple bonds between some monodentate ligands, such as the oxoligand, ═O, and the metal atom.

In one embodiment, where the linker(s) is (are) capable of directlybinding to or chelating the metal of the imaging moiety, and where boththe linker(s) and the terminal ligand(s) are monodentate and occupy onlyone valence per linker and one valence per terminal ligand, theinvention comprises compounds of the formula:

[(NSAID, NSAID residue, or NSAID derivative)-monodentatelinker]_(x)-M-(terminal monodentate ligand)_(y)

where “NSAID, NSAID residue, or NSAID derivative” refers to a NSAID or aresidue or derivative of a NSAID, M is selected from the groupconsisting of ^(99m)Tc, ⁵²Mn, and Re, x is an integer between 1 and 6inclusive, y is an integer between 0 and 5 inclusive, and x+y=6, and“terminal ligand” is a ligand that coordinates to ^(99m)Tc, ⁵²Mn, or Re,but which does not have a NSAID or a residue or derivative of a NSAIDattached to it.

Chelating groups (including linkers, where the linker is capable ofchelation) and terminal ligands may be monodentate or polydentate, withthe condition that the chelating group or groups and terminal ligand orligands must together satisfy the valency of the metal atom to whichthey are bonded. For example, in the following compound:

the mertiatide moiety is tetradentate and occupies four valences of therhenium ion, while the monodentate oxo ligand occupies two valences ofthe rhenium ion, thus together satisfying the valency of the Re(VI) ion.

In the formula:

(NSAID, NSAID derivative, or NSAID residue)-(linker)-(chelatinggroup)-M-(terminal ligand)_(z2),

which can be written as the three individual formulas:

(NSAID)-(linker)-(chelating group)-M-(terminal ligand)_(z2)

(NSAID derivative)-(linker)-(chelating group)-M-(terminal ligand)_(z2)

(NSAID residue)-(linker)-(chelating group)-M-(terminal ligand)_(z2),

and where z2 is an integer between 0 and 4 inclusive,the chelating group binds the radioactive agent M by two or more bonds.When two or more terminal ligands are present, two or more of theterminal ligands can be combined to form an additional chelating groupor groups.

Examples of terminal ligands include: halo (—X), fluoro (—F), chloro(—Cl), bromo (—Br), iodo (—I), oxo (═O), cyano (—CN), isocyano (—NC),hydroxyl (—OH), with oxo being a preferred terminal ligand; andsubstituted isocyano ligands of the form:

where R¹, R², and R³ can be independently selected from the groupconsisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted withhydroxy, fluoro, C₁-C₄ alkyl, or C₃-C₆ cycloalkyl, or R¹ and R² togetherwith the carbon to which they are attached form a C₃-C₈ cycloalkyl ringor heterocycloalkyl ring, such as

These substituted isocyano ligands are preferred when the NSAID linkeris also an isocyano ligand.

Examples of cyclooxygenase inhibitors with a linker which incorporates aligand to bind ^(99m)Tc, which can be used as the moiety [(NSAID, NSAIDresidue, or NSAID derivative)-linker] include, but are not limited to:

where the encircled portion represents a residue of ketorolac (where oneof the hydrogens on the phenyl ring of ketorolac has been replaced bythe linker);where the encircled portion represents a residue of naproxen (where themethyl of the methoxy group of naproxen has been replaced by thelinker);where the encircled portion represents a residue of ibuprofen (where oneof the methyl groups of the (2-methylpropyl) group of ibuprofen has beenreplaced by the linker); andwhere the encircled portion represents a residue of indomethacin (wherethe methyl of the methoxy group of indomethacin has been replaced by thelinker). The linkers are attached as indicated above in order topreserve the pharmacophoric portions of the NSAID.

In some embodiments, when x is two or greater, all of the NSAID, NSAIDresidue, or NSAID derivatives in the x subunits [(NSAID, NSAID residue,or NSAID derivative)-linker] are the same NSAID, NSAID residue, or NSAIDderivative. In some embodiments, when x is two or greater, at least oneof the NSAID, NSAID residue, or NSAID derivatives in the x subunits of[(NSAID, NSAID residue, or NSAID derivative)-linker] is different thanthe remaining NSAID(s) or NSAID residue(s) in the remaining subunits of[(NSAID, NSAID residue, or NSAID derivative)-linker]. In someembodiments, when x is two or greater, each of the NSAID, NSAID residue,or NSAID derivatives in the x subunits of [(NSAID, NSAID residue, orNSAID derivative)-linker] is different than the remaining NSAID(s) orNSAID residue(s) in the other subunits of [(NSAID, NSAID residue, orNSAID derivative)-linker].

Exemplary conjugates of the invention can be of the form:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, where R⁸ is optionally substituted C₁-C₄ alkyl, R⁴ isselected from the group consisting of optionally substituted C₁-C₃₀hydrocarbylene and optionally substituted C₂-C₃₀ heterohydrocarbylenegroup (such as C₁-C₁₂ alkylene), C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀ aryl-C₁-C₆ alkyl; L_(F) isabsent or is a functional group selected from the group consisting of—(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—,—N(H)—(C═O)—, —(SO₂) N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹) (C═O)N(R⁹)—,—N(R⁹)—(C═O)—O—, and —O—(C═O)N(R⁹)— where one valence of L_(F) (whenL_(F) is present) is attached to the R⁴ group and the other valence isattached to the NSAID or residue or derivative of a NSAID; and whereeach R⁹ is independently selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl (in one embodiment, R⁹ is H), and Mis selected from the group consisting of ^(99m)Tc, ⁵²Mn, and Re, whereRe can be ¹⁸⁶Re or ¹⁸⁸Re.

Examples of specific conjugates of this form include:

Further exemplary conjugates of the invention can be of the form:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, where R⁸ is optionally substituted C₁-C₄ alkyl, R⁴ isselected from the group consisting of optionally substituted C₁-C₃₀hydrocarbylene, optionally substituted C₂-C₃₀ heterohydrocarbylene group(such as C₁-C₁₂ alkylene), C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀ aryl-C₁-C₆ alkyl; R⁵ is —OH,—NH₂, —NH(C₁-C₄ alkyl), or —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), L_(E) is absentor is a functional group selected from the group consistingof —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—,—N(H)—(C═O)—, —(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹) (C═O)N(R⁹)—,—N(R⁹)—(C═O)—O—, and —O—(C═O)N(R⁹)— where one valence of L_(F) (whenL_(F) is present) is attached to the R⁴ group and the other valence isattached to the NSAID or residue or derivative of a NSAID; and whereeach R⁹ is independently selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl (in one embodiment, R⁹ is H); and Mis selected from the group consisting of ⁹⁹Tc, ⁵²Mn, and Re, where Recan be ¹⁸⁶Re or ¹⁸⁸Re. In a further embodiment, R⁵ is selected from thegroup consisting of —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄alkyl), —OCH₃, and —OCH₂CH₃; in yet a further embodiment, R⁵ is selectedfrom the group consisting of —NH₂, —OCH₃, and —OCH₂CH₃.

Examples of specific conjugates of this form include:

Further exemplary conjugates of the invention can be of the form:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, where R⁸ is optionally substituted C₁-C₄ alkyl, R⁴ isselected from the group consisting of optionally substituted C₁-C₃₀hydrocarbylene, optionally substituted C₂-C₃₀ heterohydrocarbylene group(such as C₁-C₁₂ alkylene), C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀ aryl-C₁-C₆ alkyl; R⁵ is —OH,—NH₂, —NH(C₁-C₄ alkyl), or —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), L_(F) is absentor is a functional group selected from the group consisting of —(C═O)—,—O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—,—(SO₂) N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹) (C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, and—O—(C═O)N(R⁹)— where one valence of L_(F) (when L_(F) is present) isattached to the R⁴ group and the other valence is attached to the NSAIDor residue or derivative of a NSAID; and where each R⁹ is independentlyselected from the group consisting of H and optionally substituted C₁-C₄alkyl (in one embodiment, R⁹ is H); R_((n)) is —CH₃ or —CH₂CH₃ and p isan integer between 0 and 4 inclusive; and M is selected from the groupconsisting of ^(99m)Tc, ⁵²Mn, and Re, where Re can be ¹⁸⁶Re or ¹⁸⁸Re. Ina further embodiment, R⁵ is selected from the group consisting of —NH₂,—NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —OCH₃, and —OCH₂CH₃; inyet a further embodiment, R⁵ is selected from the group consisting of—NH₂, —OCH₃, and —OCH₂CH₃

Examples of specific conjugates of this form include:

Further exemplary conjugates of the invention can be of the form:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, where R⁸ is optionally substituted C₁-C₄ alkyl, R⁴ isselected from the group consisting of optionally substituted C₁-C₃₀hydrocarbylene, optionally substituted C₂-C₃₀ heterohydrocarbylene group(such as C₁-C₁₂ alkylene), C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀ aryl-C₁-C₆ alkyl; R⁵ is —OH,—NH₂, —NH(C₁-C₄ alkyl), or —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), L_(F) is absentor is a functional group selected from the group consisting of —(C═O)—,—O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹) (C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, and—O—(C═O)N(R⁹)— where one valence of L_(F) (when L_(F) is present) isattached to the R⁴ group and the other valence is attached to the NSAIDor residue or derivative of a NSAID; and where each R⁹ is independentlyselected from the group consisting of H and optionally substituted C₁-C₄alkyl (in one embodiment, R⁹ is H); and M is selected from the groupconsisting of ⁹⁹Tc, ⁵²Mn, and Re, where Re can be ¹⁸⁶Re or ¹⁸⁸Re. In afurther embodiment, R⁵ is selected from the group consisting of —NH₂,—NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —OCH₃, and —OCH₂CH₃; inyet a further embodiment, R⁵ is selected from the group consisting of—NH₂, —OCH₃, and —OCH₂CH₃.

Examples of specific conjugates of this form include:

Further exemplary conjugates of the invention can be of the form:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, where R⁸ is optionally substituted C₁-C₄ alkyl, R⁴ isselected from the group consisting of optionally substituted C₁-C₃₀hydrocarbylene, optionally substituted C₂-C₃₀ heterohydrocarbylene group(such as C₁-C₁₂ alkylene), C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀aryl-C₁-C₆ alkyl, and C₁-C₆ aryl-C₁-C₆ alkyl; R⁵ is —OH, —NH₂, —NH(C₁-C₄alkyl), or —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), L_(F) is absent or is afunctional group selected from the group consistingof —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —(C═O)N(H)—, —N(H)—(C═O)—,—N(R⁹)—(C═O)—, —(SO₂) N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹) (C═O)N(R⁹)—,—N(R⁹)—(C═O)—O—, and —O—(C═O)N(R⁹)— where one valence of L_(F) (whenL_(F) is present) is attached to the R⁴ group and the other valence isattached to the NSAID or residue or derivative of a NSAID; and whereeach R⁹ is independently selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl (in one embodiment, R⁹ is H); and Mis selected from the group consisting of ⁹⁹Tc, ⁵²Mn, and Re, where Recan be ¹⁸⁶Re or ¹⁸⁸Re. In a further embodiment, R⁵ is selected from thegroup consisting of —NH₂, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄alkyl), —OCH₃, and —OCH₂CH₃; in yet a further embodiment, R⁵ is selectedfrom the group consisting of —NH₂, —OCH₃, and —OCH₂CH₃

Examples of specific conjugates of this form include:

All pharmaceutically acceptable salts of all of the above general andspecific compounds are also included in this disclosure.

Specific exemplary technetium conjugates of the invention are disclosedin the Examples and in FIG. 1. Specific exemplary rhenium conjugates ofthe invention are disclosed in the Examples and in FIG. 2. Furtherspecific conjugates of the invention are disclosed in the Examples andin FIG. 3.

Formulations and Routes of Administration

The compounds of the invention, that is, any conjugates of the inventiondisclosed herein, used in the methods of the invention may beadministered in any suitable form that will provide sufficient levels ofthe compounds for the purposes of imaging. Intravenous administration isa useful route of administration, although other parenteral routes canalso be employed, where parenteral as used herein includes subcutaneousinjections, intravenous injection, intraarterial injection,intramuscular injection, intrasternal injection, intraperitonealinjection, or infusion techniques. The compounds can also beadministered orally or enterally, which is a preferred route whencompatible with the absorption of the compound and with imagingrequirements. Where the pharmacokinetics of the compounds are suitable,the compounds can also be administered sublingually, by buccaladministration, subcutaneously, by spinal administration, by epiduraladministration, by administration to cerebral ventricles, by inhalation(e.g. as mists or sprays), rectally, or topically in unit dosageformulations containing conventional nontoxic pharmaceuticallyacceptable carriers, excipients, adjuvants, and vehicles as desired. Thecompounds may be administered directly to a specific or affected organor tissue. The compounds are mixed with pharmaceutically acceptablecarriers, excipients, adjuvants, and vehicles appropriate for thedesired route of administration.

In certain embodiments of the invention, especially those embodimentswhere a formulation is used for injection or other parenteraladministration, including the routes listed herein, but also includingany other route of administration described herein (such as oral,enteric, gastric, etc.), the formulations and preparations used in themethods of the invention are sterile. Sterile pharmaceuticalformulations are compounded or manufactured according topharmaceutical-grade sterilization standards (United States PharmacopeiaChapters 797, 1072, and 1211; California Business & Professions Code4127.7; 16 California Code of Regulations 1751, 21 Code of FederalRegulations 211) known to those of skill in the art.

Oral administration is advantageous due to its ease of implementationand patient compliance. If a patient has difficulty swallowing,introduction of medicine via feeding tube, feeding syringe, orgastrostomy can be employed in order to accomplish entericadministration. The active compound (and, if present, otherco-administered agents) can be enterally administered in any otherpharmaceutically acceptable carrier suitable for formulation foradministration via feeding tube, feeding syringe, or gastrostomy.

Intravenous administration can also be used advantageously, for deliveryof the conjugates of the invention to the bloodstream as quickly aspossible and to circumvent the need for absorption from thegastrointestinal tract.

The compounds described for use herein can be administered in solidform, in liquid form, in aerosol form, or in the form of tablets, pills,powder mixtures, capsules, granules, injectables, solutions,suppositories, enemas, colonic irrigations, emulsions, dispersions, foodpremixes, and in other forms suitable for the route of administration.The compounds can also be administered in liposome formulations. Thecompounds can also be administered as prodrugs, where the prodrugundergoes transformation in the treated subject to a therapeuticallyeffective form. Additional methods of administration are known in theart.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to methods known inthe art using suitable dispersing or wetting agents and suspendingagents. The sterile injectable preparation may also be a sterileinjectable solution or suspension in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in propylene glycol.Among the acceptable vehicles and solvents that may be employed arewater, Ringer's solution, and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono or di-glycerides. In addition, fattyacids such as oleic acid find use in the preparation of injectables.

Solid dosage forms for oral administration may include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive compound may be admixed with at least one inert diluent such assucrose, lactose, or starch. Such dosage forms may also compriseadditional substances other than inert diluents, e.g., lubricatingagents such as magnesium stearate. In the case of capsules, tablets, andpills, the dosage forms may also comprise buffering agents. Tablets andpills can additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions may also comprise adjuvants, such as wetting agents,emulsifying and suspending agents, cyclodextrins, and sweetening,flavoring, and perfuming agents. Alternatively, the compound may also beadministered in neat form if suitable.

The compounds for use in the present invention can also be administeredin the form of liposomes. As is known in the art, liposomes aregenerally derived from phospholipids or other lipid substances.Liposomes are formed by mono or multilamellar hydrated liquid crystalsthat are dispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound for use in the present invention, stabilizers,preservatives, excipients, and the like. The preferred lipids are thephospholipids and phosphatidyl cholines (lecithins), both natural andsynthetic. Methods to form liposomes are known in the art. See, forexample, Prescott, Ed., Methods in Cell Biology, Volume XIV, AcademicPress, New York, N.W., p. 33 et seq (1976).

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form can vary depending upon thepatient to which the active ingredient is administered and theparticular mode of administration. It will be understood, however, thatthe specific dose level for any particular patient will depend upon avariety of factors including the specific compound employed; the age,body weight, body area, body mass index (BMI), general health, sex, anddiet of the patient; the time of administration and route ofadministration used; the rate of excretion; and the drug combination, ifany, used. The compounds can be administered in a unit dosageformulation. The pharmaceutical unit dosage chosen is fabricated andadministered to provide sufficient concentration of drug for imaging apatient.

While the compounds for use in the present invention can be administeredas the sole active pharmaceutical agent, they can also be used incombination with one or more other agents. When additional active agentsare used in combination with the compounds for use in the presentinvention, the additional active agents may generally be employed intherapeutic amounts as indicated in the Physicians' Desk Reference (PDR)53rd Edition (1999), which is incorporated herein by reference, or suchtherapeutically useful amounts as would be known to one of ordinaryskill in the art, or as are determined empirically for each patient.

Combinations of the NSAID conjugates can also be used. Combining two ormore conjugates of different NSAIDs or residues or derivative s ofNSAIDs can provide advantages over using a single conjugate. Advantagescan include the ability to tune pharmacokinetics and pharmacodynamics,to adjust the solubility of the overall composition and/or itscomponents, to adjust the half-life of total conjugate in the body, toenhance imaging contrast and/or definition, to adjust binding kineticsto COX, to adjust binding affinity to COX, or to enhance the stabilityof the composition either in storage or in use. The two or moreconjugates can be combined in solution form such as those solution formsdescribed above (such as in a sterile solution for IV administration),or in solid form such as those solid forms as described above (such aspill or tablet form). The two or more conjugates can be mixed togethershortly before administration and administered together. The two or moreconjugates can be administered simultaneously, either by the same routeof administration or by different routes of administration. The two ormore conjugates can be administered consecutively, either by the sameroute of administration or by different routes of administration. In oneembodiment, a kit form can contain two or more conjugates as individualconjugates, with printed or electronic instructions for administrationeither as a mixture of conjugates, as separate conjugates administeredsimultaneously, or as separate conjugates administered consecutively.Where three or more conjugates are administered, they can beadministered as a mixture of conjugates, as separate conjugatesadministered simultaneously, as separate conjugates administeredconsecutively, as separate conjugates where two or more may beadministered simultaneously with the remainder administeredconsecutively before or after the simultaneous administration, or anyother possible combination of mixed administration, simultaneousadministration, and consecutive administration.

Imaging Techniques

The conjugates can be used with any suitable imaging technique. Imagesof a subject, or of a portion of a subject such as the arm, leg, or anyspecific region of the body of the subject, can be generated using gammacameras, planar gamma imaging, scintigraphic imaging, SPECT imaging(single photon emission computed tomography), and other radiographic ortomographic imaging techniques. Exemplary imaging methods that can beused are described in Pacelli et al., J. Label. Compd. Radiopharm.57:317-322 (2014); de Vries et al., J Nucl. Med. 44:1700-1706 (2003);Tietz et al., Current Medicinal Chemistry, 20, 4350-4369 (2013); Sogbeinet al., Biomed. Res. Int., Sogbein, Oyebola O. et al., BioMed ResearchInternational, 2014:942960, doi: 10.1155/2014/942960; and Wernick, M. N.and Aarsvold, J. N., Emission Tomography: The Fundamentals of PET andSPECT, San Diego: Elsevier Academic Press, 2004.

Kits

Further embodiments of the invention embrace one or more kit forms whichcan contain one or more conjugates as disclosed herein. The kit cancontain printed or electronic instructions for administration of theconjugate. In further embodiments, the kit can contain one or morecompounds as disclosed herein which lacks the radioactive agent, withprinted or electronic instructions for adding the radioactive agent toconstitute one or more conjugates of the invention.

The following examples are intended to illustrate, but not limit, theinvention.

EXAMPLES Synthetic Examples Example 1 General Method for the synthesisoftetrakis(((2-((6-((S)-1-carboxyethyl)naphthalen-2-yl)oxy)-2-methylpropyl)-14-azanylidyne)methyl)copper (2)

Synthesis of (S)-2-(6-hydroxynaphthalen-2-yl)propanoic acid (4)

As described in del Amo, V., McGlone, A. P., Soriano, J. M. & Davis, A.P. “Two-colour screening in combinatorial chemistry: prospecting forenantioselectivity in a library of steroid-based receptors.” Tetrahedron65, 6370-6381 (2009), (S)-2-(6-methoxynaphthalen-2-yl)propanoic acid(Naproxen) (3) (3.0 g, 13.0 mmol) is suspended in HBr (48% w/w in water,120 mL) and heated at reflux for 2 h. The mixture is allowed to warm toroom temperature and filtered. The solid is washed with chilled water toafford the title product (S)-2-(6-hydroxynaphthalen-2-yl)propanoic acid(4).

Synthesis of tert-butyl (S)-2-(6-hydroxynaphthalen-2-yl)propanoate (5)

As described in del Amo, V., McGlone, A. P., Soriano, J. M. & Davis, A.P. “Two-colour screening in combinatorial chemistry: prospecting forenantioselectivity in a library of steroid-based receptors.” Tetrahedron65, 6370-6381 (2009), the carboxylic acid (4) (2.39 g, 11.05 mmol) isdissolved in dry THF (110 mL) under a nitrogen atmosphere. The solutionis cooled to 0° C., then trifluoroacetic anhydride (13.93 g, 9.42 mL,66.32 mmol) is added drop wise and the mixture is further stirred for 4h maintaining the temperature below 5° C. tert-Butanol (32 mL) is addeddrop wise and the resulting mixture is then allowed to rise to roomtemperature and is vigorously stirred overnight. The reaction is cooledagain to ice-bath temperature and NH₄OH (35% in water, 6 mL) is addeddrop wise. When the addition is finished the mixture is allowed to riseto room temperature again and is finally stirred for a further 30 minbefore the volatiles are evaporated under vacuum. The crude residue istriturated with boiling DCM and the crystalline solid formed is removedby filtration. The filtrate is washed with saturated aqueous NaHCO₃ anddried over MgSO₄, then the solvent is evaporated under reduced pressureto give the tert-butyl ester (5). An analytical sample can be preparedby crystallizing (5) from hot benzene/hexane.

Synthesis of tert-butyl(S)-2-(6-((1-formamido-2-methylpropan-2-yl)oxy)naphthalen-2-yl)propanoate(6)

Using the method of Al-Ktaifani, M. M., Nakawa, A. a., Tabbaa, Z. a. &Namou, A. a. “Synthesis of 2-methyl-2-propoxypropyl isocyanide and itsCu(I) tetrafluoroborate complex.” Chem. Pap. 62, 329-333 (2008): ingeneral, mercuric(II)trifluoroacetate andN-(2-methyl-2-propenyl)formamide (Synthesized by the method of van Wyk,A. J. et al. “Synthesis and 99mTc labelling of MMI (MIBI) and its ethylanalogue EMI.” Int. J. Radiat. Appl. Instrumentation. Part A. Appl.Radiat. Isot. 42, 687-689 (1991)) and tert-butyl(S)-2-(6-hydroxynaphthalen-2-yl)propanoate (5) are dissolved in THF andstirred at room temperature. The mixture is cooled down in an ice bathand NaOH pellets in isopropanol are added slowly. Subsequently, NaBH₄was added to the mixture, which was stirred for 18 h. The solution waspassed through a column of Celite® (J. T. Baker, Phillipsberg, N.J.,diatomaceous earth) to remove elemental Hg to yield tert-butyl(S)-2-(6-((1-formamido-2-methylpropan-2-yl)oxy)naphthalen-2-yl)propanoate(6).

Synthesis of(S)-2-(6-((1-isocyano-2-methylpropan-2-yl)oxy)naphthalen-2-yl)propanoicacid (7)

tert-Butyl(S)-2-(6-((1-formamido-2-methylpropan-2-yl)oxy)naphthalen-2-yl)propanoate(6) is dissolved in pyridine and benzene. The solution is cooled to 0°C. and POCl₃ is subsequently added drop wise. The reaction is quenchedby pouring over ice and the product is extracted into CH₂Cl₂.

Synthesis oftetrakis(((2-((6-((S)-1-carboxyethyl)naphthalen-2-yl)oxy)-2-methylpropyl)-14-azanylidyne)methyl)copper (2)

Based on the method of Al-Ktaifani, M. M., Nakawa, A. a., Tabbaa, Z. a.& Namou, A. a. “Synthesis of 2-methyl-2-propoxypropyl isocyanide and itsCu(I) tetrafluoroborate complex.” Chem. Pap. 62, 329-333 (2008):synthesis of (2) is realized by heating a mixture of (7) and freshlycrystallized CuCl in degassed dry EtOH at 50° C. in an ampoule for 1 h.A solution of NaBF₄ in water is added in one portion, solvents areremoved under reduced pressure, and the resultant crude product isdissolved in EtOH and the NaCl generated is separated by filtration. Thesolvent is removed to get the product (2).

Example 2 Synthesis oftetrakis(((2-(4-(((S)-1-carboxy-2,3-dihydro-1H-pyrrolizine-5-carbonyl)phenethoxy)-2-methylpropyl)-14-azanylidyne)methyl)copper (16)

Synthesis of 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)benzoic acid (9)

Commercially available 4-(2-hydroxyethyl)benzoic acid (8) is protectedas the TBDMS ether using standard conditions (see, for example, Greene'sProtective Groups in Organic Synthesis, 4th Edition. Peter G. M. Wuts,Theodora W. Greene; John Wiley & Sons, Inc.; ISBN: 978-0-471-69754-1).

Synthesis of 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)benzoyl chloride(10)

4-(2-((tert-butyldimethylsilyl)oxy)ethyl)benzoic acid (9) is convertedto its acid chloride using standard conditions.

Synthesis of tert-butyl(S)-5-(4-(2-((tert-butyldimethylsilyl)oxy)ethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylate(12) Components (10) and tert-butyl(S)-2,3-dihydro-1H-pyrrolizine-1-carboxylate (11) are coupled using themethods of either Muchowski et al. or Baran et al. (for the chiralsynthesis) of (12) (Muchowski, J. M. et al., “Synthesis andantiinflammatory and analgesic activity of5-aroyl-1,2-dihydro-3H-pyrrolo[1,2-a]pyrrole-1-carboxylic acids andrelated compounds,” J. Med. Chem. 28, 1037-49 (1985); Baran, P. S.,Richter, J. M. and Lin, D. W., “Direct coupling of pyrroles withcarbonyl compounds: short enantioselective synthesis of (S)-ketorolac,”Angew. Chem. Int. Ed. Engl. 44, 609-12 (2005)).

Synthesis of tert-butyl(S)-5-(4-(2-hydroxyethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylate(13) Compound (12) is deprotected with TBAF using standard methods toyield (13).

tert-butyl(S)-5-(4-(2-(((1-formamido-2-methylpropan-2-yl)oxy)ethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylate(14)

Using the method of Al-Ktaifani, M. M., Nakawa, A. A., Tabbaa, Z. A. andNamou, A. A., “Synthesis of 2-methyl-2-propoxypropyl isocyanide and itsCu(I) tetrafluoroborate complex,” Chem. Pap. 62, 329-333 (2008).

In general, mercuric(II)trifluoroacetate andN-(2-methyl-2-propenyl)formamide (Synthesized by the method of van Wyk,A. J. et al. “Synthesis and 99mTc labelling of MMI (MIBI) and its ethylanalogue EMI.” Int. J. Radiat. Appl. Instrumentation. Part A. Appl.Radiat. Isot. 42, 687-689 (1991)) and tert-butyl(S)-5-(4-(2-hydroxyethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylate(13) are dissolved in THF and stirred at room temperature. The mixtureis cooled down in an ice bath and NaOH pellets in isopropanol are addedslowly. Subsequently, NaBH₄ was added to the mixture, which was stirredfor 18 h. The solution was passed through a column of Celite® (J. T.Baker, Phillipsberg, N.J., diatomaceous earth) to remove elemental Hg toyield tert-butyl(S)-5-(4-(2-((1-formamido-2-methylpropan-2-yl)oxy)ethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylate(14).

Synthesis of(S)-5-(4-(2-((1-isocyano-2-methylpropan-2-yl)oxy)ethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylicacid (15) tert-butyl(S)-5-(4-(2-((1-formamido-2-methylpropan-2-yl)oxy)ethyl)benzoyl)-2,3-dihydro-1H-pyrrolizine-1-carboxylate(14) is dissolved in pyridine and benzene. The solution is cooled to 0°C. and POCl₃ is subsequently added drop wise. The reaction is quenchedby pouring over ice and extracting the product into CH₂Cl₂.

Synthesis oftetrakis(((2-(4-((S)-1-carboxy-2,3-dihydro-1H-pyrrolizine-5-carbonyl)phenethoxy)-2-methylpropyl)-14-azanylidyne)methyl)copper (16) Based on the method ofAl-Ktaifani, M. M., Nakawa, A. A., Tabbaa, Z. A. and Namou, A. A.,“Synthesis of 2-methyl-2-propoxypropyl isocyanide and its Cu(I)tetrafluoroborate complex.” Chem. Pap. 62, 329-333 (2008). Synthesis of(16) is realized by heating a mixture of (15) and freshly crystallizedCuCl in degassed dry EtOH at 50° C. in an ampoule for 1 h. A solution ofNaBF₄ in water is added in one portion, solvents are removed undervacuum, with the resultant crude product dissolved in EtOH and the NaClgenerated was separated by filtration. The solvent was removed to getthe product (16).

Example 3 Synthesis ofhexakisa(2-(((6((S)-1-carboxyethyl)naphthalen-2-yl)oxy)-2-methylpropyl)-14-azanylidyne)methyl)technetium(I)

A vial is prepared containing a sterile, non-pyrogenic, lyophilizedmixture of:

Tetrakis(((2-(((6-((S)-1-carboxyethyl)naphthalen-2-yl)oxy)-2-methylpropyl)-14-azanylidyne)methyl)Copper (I) tetrafluoroborate—1.0 mg

Sodium Citrate Dihydrate—2.6 mg

L-Cysteine Hydrochloride Monohydrate—1.0 mg

Mannitol—20 mg

Stannous Chloride, Dihydrate, minimum (SnCl₂.2H₂O)—0.025 mg

Stannous Chloride, Dihydrate, (SnCl₂.2H₂O)—0.075 mg

Tin Chloride (stannous and stannic) Dihydrate, maximum (asSnCl₂.2H₂O)—0.086 mg.

Prior to lyophilization the pH is 5.3 to 5.9. The contents of the vialare lyophilized and stored under nitrogen.

Reconstitution with sterile, non-pyrogenic, oxidant-free SodiumPertechnetate Tc99m Injection forms the compound (1), which isadministered by intravenous injection for diagnostic use (see Scheme 2).The specific steps for reconstitution are:

Place the vial in a suitable radiation shield with a fitted radiationcap.

With a sterile shielded syringe, aseptically obtain additive-free,sterile, non-pyrogenic Sodium Pertechnetate Tc99m Injection [925-5550MBq, (25-150 mCi)] in approximately 1 to 3 mL.

Aseptically add the Sodium Pertechnetate Tc99m Injection to the vial inthe lead shield. Without withdrawing the needle, remove an equal volumeof headspace to maintain atmospheric pressure within the vial.

Shake vigorously, about 5 to 10 quick upward-downward motions.

Remove the vial from the lead shield and place upright in anappropriately shielded and contained boiling water bath, such that thevial is suspended above the bottom of the bath, and boil for 10 minutes.Timing for 10 minutes is begun as soon as the water begins to boilagain. Do not allow the boiling water to come in contact with thealuminum crimp.

Remove the vial from the water bath, place in the lead shield and allowto cool for fifteen minutes.

Using proper shielding, the vial contents should be visually inspected.Use only if the solution is clear and free of particulate matter anddiscoloration.

Assay the reaction vial using a suitable radioactivity calibrationsystem. Record the Technetium Tc99m concentration, total volume, assaytime and date, expiration time and lot number on the vial shield labeland affix the label to the shield.

Store the reaction vial containing the Technetium Tc99m (1) at 15° C. to25° C. until use; at such time the product should be asepticallywithdrawn. Technetium Tc99m (1) should be used within six hours ofpreparation. The vial contains no preservative.

The pH of the reconstituted product is 5.5 (5.0-6.0). No bacteriostaticpreservative is present. The precise structure of the technetium complex(1) is Tc99m[IBN]₆ ⁺ where IBN is(S)-2-(6-((1-isocyano-2-methylpropan-2-yl)oxy)naphthalen-2-yl)propanoicacid.

Example 4

Synthesis ofHexakis(((2-(4-((S)-1-carboxy-2,3-dihydro-1H-pyrrolizine-5-carbonyl)phenethoxy)-2-methylpropyl)-14-azanylidyne)methyl)technetium(17)

Hexakis(((2-(4-((S)-1-carboxy-2,3-dihydro-1H-pyrrolizine-5-carbonyl)phenethoxy)-2-methylpropyl)-14-azanylidyne)methyl)technetium(17) is prepared using the same procedure as in Example 3, starting fromtetrakis(((2-(4-((S)-1-carboxy-2,3-dihydro-1H-pyrrolizine-5-carbonyl)phenethoxy)-2-methylpropyl)-14-azanylidyne)methyl)copper (16).

Example 5 Synthesis of Indomethacin Derivatives

Amine 20 is readily synthesized by using protocols available in thefollowing publications: (1) Uddin, Md. Jashim, et al. BioconjugateChemistry (2013), 24(4), 712-723; (2) Zlatopolskiy, Boris D., et al.Chemical Communications (2012), 48(57), 7134-7136; (3) U.S. Pat. Appl.Publ. 2005/0002859 (2005); (4) U.S. Pat. Appl. Publ. 2007/0292352(2007); (5) Uddin, Md. Jashim, et al. Journal of Labelled Compounds andRadiopharmaceuticals (2009), 52(9), 387-393; and (6) Uddin, Md. Jashim,et al. Bioorganic & Medicinal Chemistry Letters (2010), 20(5),1787-1791.

Alternatively, amine 20 can be synthesized by the following procedure:

To a stirred solution of indomethacin 18 (2 g, 5.6 mmol) in DMF (50 mL)was added tert-butyl 4-aminobutylcarbamate (2 g, 11.2 mmol), HOBt (2.3g, 16.8 mmol), DIPEA (3.6 g, 28 mmol) and EDCI (2.1 g, 11.2 mmol, 2 eq)at rt. The resultant mixture was stirred at rt for 16 h. The mixture waswashed with 100 mL saturated LiCl solution and extracted with EtOAc(3×100 mL). The organic layers were combined, concentrated andcrystallized from n-hexane to give 19 as a yellow solid. LC-MS: m/z=428(M-100+H)⁺; H-NMR (400 MHz, CDCl₃) δ: 7.68 (d, 2H, J=8.8 Hz), 7.50 (d,2H, J=8.4 Hz), 6.90 (m, 2H), 6.71 (d, 2H, J=2.4 Hz), 5.79 (s, 1H), 4.59(s, 1H), 3.82 (s, 3H), 3.63 (s, 2H), 3.22 (m, 2H), 3.06 (m, 2H), 2.39(s, 3H), 1.41 (m, 12H).

To a stirred solution of 19 (2 g, 3.7 mmol) in DCM (20 mL), a solutionof HCl in dioxane (4M, 7.5 mL) was added dropwise. The resultantsolution was stirred at rt for 2 h. Removal of solvent under reducedpressure afforded 20 as a yellow residue, which was used without furtherpurification. LC-MS: m/z=428 (M-36+H)⁺; H-NMR (400 MHz, DMSO-d₆) δ: 8.26(s, 1H), 8.07 (s, 3H), 7.68 (m, 4H), 7.18 (d, 1H, J=2.4 Hz), 6.94 (d,2H, J=8 Hz), 6.71 (d, 1H, J=2.8 Hz), 3.77 (s, 3H), 3.56 (s, 2H), 3.07(d, 2H, J=6 Hz), 2.76 (d, 2H, J=6.4 Hz), 2.24 (s, 3H), 1.57 (m, 2H),1.48 (m, 2H).

Indomethacin derivative 22 is made following the procedure in: Wang Y,et al. Nat Protoc. (2007), 2(4): 972-8. The reagent S-Acetyl-MAG3-NHSester is available commercially (KeraFAST Inc., Boston, Mass., USA;catalog no. ES1001; see the supplier web site:www.kerafast.com/p-1447-s-acetyl-mag3-nhs-ester.aspx?gclid=Cj0KEQjwur2eBRDtvMS0gIuS-dYBEiQANBPMR3SKY-ABw0HCC08Gud53dB29wsldYYU13UeQFvVxc_gaAiIA8P8HAQ;see also Winnard P. et al., Nucl Med Biol. (1997), 24(5): 425-32; WangY. et al., Nat. Protoc. (2007) 2(4): 972-8; Rusckowski M. et al., CancerBiotherapy & Radiopharmaceuticals (2007), 22(4): 564-72; and Wang Y. etal., European J. Nucl. Med. Mol. Imag. (2009), 36(12): 1977-86).

Indomethacin derivative 23 is made following the procedure in Ono, etal. Bioorg. Med. Chem. Lett. (2010), 20, 5743-5748. The reagentTrichlorooxobis(triphenylphosphine)rhenium(V), (PPh3)2ReOCl₃, iscommercially available (Sigma-Aldrich, Saint Louis, Mo., USA; catalogno. 370193).

Alternatively, compound 23 can be synthesized using the followingprocedure: To a stirred solution of compound 22 (100 mg, 0.14 mmol) inNMP (15 mL), (PPh₃)₂ReOCl₃ (Sigma-Aldrich, Order #370193, 106 mg, 0.127mmol) was added. The mixture was stirred at 80° C. for 48 h. Thesolution was purified by Prep-HPLC (Column: Acquity BEH C18, WatersCorp, solvent A: Water (10 mM NH₄HCO₃), solvent B: MeCN) to givecompound 23 as a solid. LC-MS: m/z=871 (M+H)⁺; H-NMR (400 MHz, DMSO-d₆)δ: 8.00 (m, 1H), 7.70˜7.63 (m, 4H), 7.20 (m, 1H), 7.11 (m, 1H), 6.95 (d,1H, J=8.8 Hz), 6.71 (d, 1H, J=2 Hz), 4.82 (d, 1H, J=16 Hz), 4.55 (d, 1H,J=17.6 Hz), 4.41 (d, 1H, J=16 Hz), 3.80 (m, 4H), 3.65 (d, 1H, J=16.8Hz), 3.48 (s, 2H), 3.03 (m, 4H), 2.22 (s, 3H), 1.36 (s, 4H). ¹³C-NMR(400 MHz, DMSO-d₆) δ: 192.27, 191.61, 188.45, 170.13, 169.65, 168.32,156.01, 137.97, 135.53, 134.77, 131.61, 130.72, 129.49, 115.00, 111.76,102.30, 58.45, 56.21, 55.90, 53.08, 39.94, 38.53, 31.61, 27.12, 26.91,13.84.

Example 6 Synthesis of Indomethacin Derivatives

Similarly as Example 5, amine 25 has been reported in the followingarticles: (1) Uddin, Md. Jashim, et al. Bioconjugate Chemistry (2013),24(4), 712-723. (2) Hua Zhang, et al. J. Am. Chem. Soc. (2013), 135,11663-11669 (3) Hua Zhang, et al. J. Am. Chem. Soc. (2013), 135,17469-17475.

Alternatively, amine 25 can be synthesized by the following procedure:To a mixture of indomethacin (11.8 g, 33 mmol), tert-butyl6-aminohexylcarbamate (CAS No. 51857-17-1) (8.5 g, 39 mmol) and HATU(18.8 g, 50 mmol) in DMF (200 mL) was added TEA (10.1 g, 0.1 mol). Themixture was stirred at rt for 4 h and poured onto ice water (300 mL).The precipitate was collected by filtration and dried to give compound24 as a solid. LC-MS: m/z=456.3 (M-100+H; H-NMR (400 MHz, DMSO-d₆) δ:8.00 (br s, 1H), 7.77-7.63 (m, 4H), 7.11 (d, J=2.4 Hz, 1H), 6.95-6.93(m, 1H), 6.74-6.69 (m, 2H), 3.76 (s, 3H), 3.48 (s, 2H), 3.06-3.01 (m,2H), 2.88-2.83 (m, 2H), 2.23 (s, 3H), 1.36 (s, 9H), 1.33-1.18 (m, 8H).

A mixture of compound 24 (10 g, 18 mmol) in HCl/dioxane (3M, 150 mL) wasstirred at rt for 2 h. The mixture was concentrated to give compound 25as white solid, which was in the next step without further purification.LC-MS: m/z=456.3 (M+H)⁺; H NMR (400 MHz, CD3OD) δ: 7.72 (d, J=8.4 Hz,2H), 7.59 (d, J=8.4 Hz, 2H), 7.03 (d, J=2.4 Hz, 1H), 6.95 (d, J=9.2 Hz,1H), 6.73-6.70 (m, 1H), 3.83 (s, 3H), 3.63 (s, 2H), 3.34-3.23 (m, 2H),2.91-2.85 (m, 2H), 2.34 (s, 3H), 1.62-1.37 (m, 8H).

Indomethacin derivative 26 is made following the procedure in: Wang Y,et al. Nat Protoc. (2007), 2(4): 972-8. The reagent S-Acetyl-MAG3-NHSester is available commercially (KeraFAST Inc., Boston, Mass., USA;catalog no. ES1001; see the supplier web site:www.kerafast.com/p-1447-s-acetyl-mag3-nhs-ester.aspx?gclid=Cj0KEQjwur2eBRDtvMS0gIuS-dYBEiQANBPMR3SKY-ABw0HCC08Gud53dB29wsldYYU13UeQFvVxc_gaAiIA8P8HAQ;see also Winnard P. et al., Nucl. Med Biol. (1997), 24(5): 425-32; WangY. et al., Nat. Protoc. (2007), 2(4): 972-8; Rusckowski M. et al.,Cancer Biotherapy & Radiopharmaceuticals (2007), 22(4): 564-72; and WangY. et al., European J. Nucl. Med. Mol. Imag.; (2009), 36(12): 1977-86).

Indomethacin derivative 27 is made following the procedure in: Ono, etal. Bioorg. Med. Chem. Lett. (2010), 20, 5743-5748. The reagentTrichlorooxobis(triphenylphosphine)rhenium(V) is commercially available(Sigma-Aldrich, Saint Louis, Mo., USA; catalog no. 370193).

Alternatively, compound 27 can be synthesized using the followingprocedure: A mixture of compound 26 (200 mg, 0.27 mmol) and(Ph₃P)₂ReOCl₃ (337 mg, 0.41 mmol) in NMP (10 mL) was stirred for 16 h at100° C. The solution was directly purified by Prep-HPLC (Column: AcquityBEH C18, Waters Corp, solvent A: water containing 10 mM NH₄HCO₃, solventB: ACN) to give 27 as a solid. LC-MS: m/z=899.2 MS (M−H)⁻; H-NMR (400MHz, DMSO-d₆) δ: 8.00 (s, 1H), 7.70-7.63 (m, 4H), 7.16-7.11 (m, 2H),6.94 (d, J=8.8 Hz, 1H), 6.71-6.69 (m, 1H), 4.78 (d, J=16.4 Hz, 1H), 4.52(d, J=18.0 Hz, 1H), 4.38 (d, J=16.0 Hz, 1H), 4.16-4.10 (m, 3H),3.79-3.75 (m, 4H), 3.63 (d, J=17.2 Hz, 1H), 3.47 (s, 2H), 3.03-2.98 (m,4H), 2.22 (s, 3H), 1.36-1.20 (m, 8H).

Example 7 Synthesis of Indomethacin Derivatives

Compound 31 has been reported by Yoriko Iwata, et al. J. Med. Chem.(2001), 44, 1718-1728 and is readily synthesized from the commerciallyavailable hydrazine 28 through the Fisher indole synthesis withlevulinic acid. Alkylation of 31 with ^(t)BocNH(CH₂)₄Br and deprotectionof the ^(t)BOC amine and the ^(t)Bu-ester leads to amino acid 33 whichcan be reacted with NHS—S-Acetyl-MAG3 following the procedure in: WangY, et al. Nat Protoc. 2007; 2(4), 972-8 to afford 34. The reagentS-Acetyl-MAG3-NHS ester is available commercially (KeraFAST Inc.,Boston, Mass., USA; catalog no. ES1001; see the supplier web site:www.kerafast.com/p-1447-s-acetyl-mag3-nhs-ester.aspx?gclid=Cj0KEQjwur2eBRDtvMS0gIuS-dYBEiQANBPMR3S KY-ABw0HCC08Gud53 dB29wsldYYU13UeQFvVxc_gaAiIA8P8HAQ;see also Winnard P. et al., Nucl. Med. Biol. (1997), 24(5): 425-32; WangY. et al., Nat. Protoc. (2007), 2(4): 972-8; Rusckowski M. et al.,Cancer Biotherapy & Radiopharmaceuticals (2007), 22(4): 564-72; and WangY. et al., European J. Nucl. Med. Mol. Imag. (2009), 36(12): 1977-86).

The rhenium-indomethacin derivative 35 is made following the procedurein: Ono, et al. Bioorg. Med. Chem. Lett. (2010), 20, 5743-5748. Thereagent trichlorooxobis(triphenylphosphine)rhenium(V) is commerciallyavailable (Sigma-Aldrich, Saint Louis, Mo., USA; catalog no. 370193).

Example 7b Synthesis of Indomethacin Derivatives

Compound 39 is prepared by a procedure similar to the one described inExample 7, substituting ^(t)BocNH(CH₂)₄Br with ^(t)BocNH(CH₂)₆Br.

Example 8 Synthesis of Ethylenedicysteine Indomethacin Derivatives 40,41, 42, and 43

Part 1: Synthesis of Ethylenedicysteine Indomethacin Derivatives 40, 41,42, and 43

Amine 20 is prepared as described in Example 5.

Indomethacin derivative 40 is made from amine 20 by amide coupling withethylenedicysteine (CAS #14344-48-0). The reaction of ethylenedicysteinewith primary amines is described in the literature, see e.g. Yang etal., US 2004/6692724.

Derivative 41 is made by reacting 40 with diazomethane orTMS-diazomethane. Analog 42 is made by treating 40 with EtOH/HCl orEtOH/TMSCl. Derivative 43 is made from 40 by reaction with NH₃ in thepresence of a dehydrating agent or by a similar method well known to aperson skilled in the art.

Example 8b Synthesis of Isocyano Indomethacin Derivative 45

The isocyano compound 45 is prepared by formylating amine 20 to yield44, followed by dehydration of 44 with POCl₃ to give 45.

Example 8c Synthesis of ^(99m)Tc Indomethacin Derivative 50b

Compound 50b is prepared from compound 41 in a manner analogous to theprocedure in Example 2.

Example 9 Synthesis of ^(99m)Tc Indomethacin Derivatives: 46, 47, 48,and 49

Compounds 46, 47, 48, and 49 are prepared from compounds 42, 43, 44, and45, respectively, by treating with sodium pertechnetate as described inExample 3.

Example 10 Synthesis of ^(99m)Tc Indomethacin MAG3 Analog (51)

Compound 51 is prepared by combining, in a reaction vial, 1 mg of 22,between 0.05 mg (minimum) stannous chloride dihydrate (SnCl₂.2H₂O) and0.2 mg (maximum) total tin expressed as stannous chloride dihydrate(SnCl₂.2H₂O), 40 mg sodium tartrate dehydrate (Na₂C₄H₂O₆.2H₂O), and 20mg lactose monohydrate. The pH of the reconstituted drug is between 5.0and 6.0. A solubilizing agent and a bacteriostatic agent are optionallyused depending on the need.

The preparation of compound 51 is analogous to the preparation ofTechnetium Tc 99m Mertiatide using the TechneScan MAG3™ Kit. Details ofthe preparation are at URLwww.mallinckrodt.com/WorkArea/DownloadAsset.aspx?id=616.

Example 11 Synthesis of ^(99m)Tc Indomethacin MAG3 Analog (52)

Compound 52 is prepared from compound 34 using a procedure similar tothe one described in Example 10.

Example 11b Synthesis of ^(99m)Tc Indomethacin MAG3 Analog (53)

Compound 53 is prepared from compound 34 using a procedure similar tothe one described in Example 10.

Example 12 Synthesis of ^(99m)Tc Indomethacin MAG3 Analog (54)

Compound 54 is prepared from compound 26 using a procedure similar tothe one described in Example 10.

Example 13 Synthesis of Ketorolac MAG3 Ligand Part 1

Compound 61 is prepared from compound 55 by first protecting the acidfunctionality of 55 via acid-catalyzed esterification to give 56, andthen using a procedure analogous to that given in Example 7.

Example 14 Synthesis of Ketorolac Hexanoic MAG Re Ligand

Compound 61 is prepared from compound 55 by first protecting the acidfunctionality of 55 via acid-catalyzed esterification to give 56, andthen using a procedure analogous to that given in Example 7.

Example 15 Synthesis of Ketorolac Ethylenedicysteine Ligand Part 1

Compound 67 is prepared from compound 55 by preparation of 57 via aprocedure analogous to that given in Example 7 using N-Boc-protected1-amino-4-bromobutane, and then by following the deprotection andcoupling steps given in Example 8.

Part 2

Compound 70 is prepared from compound 68 in a similar manner to Part 1of this Example.

Part 3

Compound 72 is prepared from compound 58 in a manner similar to Example8, Part 2.

Example 16 Synthesis of Ketorolac ^(99m)Tc Complex Part 1

Compounds 73 and 74 are prepared from compounds 67 and 70, respectively,by treating with sodium pertechnetate as described in Example 3,substituting 67 and 70, respectively, for compound 2.

Part 2

Compound 76 is prepared from compound 72 using procedures analogous tothose of Example 2 and Example 3.

Example 17 Synthesis of ^(99m)Tc Ketorolac MAG3 Analog (77)

Compound 77 is prepared from compound 60 using the procedure of Example10.

Example 18 Synthesis of ^(99m)Tc Ketorolac MAG3 Analog (78)

Compound 78 is prepared from compound 65 using the procedure of Example10.

Example 19 Synthesis of Naproxen Butanyl MAG3 Ligand

Compound 79 is prepared from compound 3 using N¹-Boc-1,4-diaminobutane.

Compound 82 is prepared from compound 79 by using a procedure analogousto the corresponding steps of Example 7.

Example 20 Synthesis of Naproxen Hexanyl Ligand

Compound 86 is prepared from compound 3 using the procedure of Example19.

Example 21 Synthesis of Naproxen Derivatives

Compound 5 is obtained from the commercial starting material 4 in 82%yield as reported in WO 2013/025790 A2. Alkylation of 5 with^(t)BocNH(CH₂)₄Br and deprotection of the ^(t)BOC amine and the^(t)Bu-ester provides amino acid 88 which can be reacted withNHS—S-Acetyl-MAG3 following the procedure in: Wang Y, et al. Nat Protoc.2007; 2(4), 972-8 to afford 89. The reagent S-Acetyl-MAG3-NHS ester isavailable commercially (KeraFAST Inc., Boston, Mass., USA; catalog no.ES1001; see the supplier web site:www.kerafast.com/p-1447-s-acetyl-mag3-nhs-ester.aspx?gclid=Cj0KEQjwur2eBRDtvMS0gIuS-dYBEiQANBPMR3SKY-ABw0HCC08Gud53dB29wsldYYU13UeQFvVxc_gaAiIA8P8HAQ;see also Winnard P. et al., Nucl Med Biol. 1997 July; 24(5):425-32; WangY. et al., Nat. Protoc. 2007; 2(4):972-8; Rusckowski M. et al., CancerBiotherapy & Radiopharmaceuticals 2007; 22(4): 564-72; and Wang Y. etal., European J Nucl Med Mol Imag 2009; 36(12): 1977-86).

The rhenium-indomethacin derivative 90 is made following a proceduresimilar to the one described by Ono, et al. Bioorg. Med. Chem. Lett.(2010), 20, 5743-5748. The reagentTrichlorooxobis(triphenylphosphine)rhenium(V) is commercially available(Sigma-Aldrich, Saint Louis, Mo., USA; catalog no. 370193).

Example 22 Synthesis of ^(99m)Tc Naproxen Butanyl MAG3 Analog (91)

Compound 91 is prepared from compound 81 using the procedure of Example10.

Example 23 Synthesis of ^(99m)Tc Naproxen Hexanyl MAG3 Analog (92)

Compound 92 is prepared from compound 85 using the procedure of Example10.

Example 24 Synthesis of ^(99m)Tc Naproxen MAG3 Analog (93)

Compound 93 is prepared from compound 89 using the procedure of Example10.

Example 25 Synthesis of Celecoxib Hexanyl MAG3 Ligand

The celecoxib analog 95, in which a methyl ester replaces thetrifluoromethyl group of celecoxib, is synthesized from 94 using theprocedure described in Lill et al., Tetrahedron Letters 54, 6682-6686,(2013). The ester is hydrolyzed to give 96, and coupled withN¹-Boc-1,6-diaminohexane to yield 97. The corresponding steps of Example7 are then followed to yield 100.

Example 26 Synthesis of Celecoxib Butanyl MAG3 Ligand

Compound 104 is produced as in Example 25, usingN¹-Boc-1,4-diaminobutane in place of N¹-Boc-1,6-diaminohexane.

Example 27 Synthesis of Celecoxib Decanyl MAG3 Ligand

Compound 108 is produced as in Example 25, usingN¹-Boc-1,10-diaminodecane in place of N¹-Boc-1,6-diaminohexane.

Example 28 Synthesis of ^(99m)Tc Celecoxib MAG3 Analog (109)

Compound 109 is prepared from compound 103 using the procedure ofExample 10.

Example 29 Synthesis of ^(99m)Tc Celecoxib MAG3 Analog (110)

Compound 110 is prepared from compound 107 using the procedure ofExample 10.

Example 30 Synthesis of ^(99m)Tc Celecoxib MAG3 analog (111)

Compound 111 is prepared from compound 99 using the procedure of Example10.

Example 31 Synthesis of Compound 178

Compound 178 was prepared by the following procedure:S,S-ethylenedicysteine 172 (CAS#14344-48-0) was prepared according tothe procedures described in the literature, see e.g. Blondeau, Berse,Gravel, Can. J. Chem. 45, 49-52, (1967); Ratner, Clarke, J. Am. Chem.Soc. 59, 200-206, (1937). Compound 176 was prepared by a proceduresimilar to the one described for compounds 20 and 25 in examples 5 and6. 176 is coupled with acid 173 to give the amide 177 as described inexample 5. Compound 177 is dissolved in ethanol and treated with 5eq ofTMSCl at 40° C. for 20 h. Solvent is removed and the product is purifiedby HPLC to give the target compound 178.

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.3 Hz, 2H), 7.49 (d, J=8.3 Hz,2H), 7.00-6.80 (m, 1H), 6.84-6.47 (m, 2H), 6.16 (s, 1H), 5.90 (s, 1H),4.67 (d, J=7.1 Hz, 1H), 4.44-4.26 (m, 2H), 3.96-3.76 (m, 5H), 3.65 (s,2H), 3.61-3.26 (m, 13H), 3.23-3.05 (m, 2H), 3.05-2.83 (m, 2H), 2.38 (s,3H), 1.34 (t, J=7.1 Hz, 3H). MS (ESI) m/z: 966.1 (M+H⁺).

The following compounds 179-185 can also be prepared according to theprocedure given in Example 31 by replacing tert-butyl2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate 174 with the appropriatereagent shown in Table 1.

TABLE 1 Compound Y = Reagent used 179

tert-butyl (2-aminoethyl)carbamate 180

tert-butyl (2-(2-aminoethoxy)ethyl)carbamate 181

tert-butyl (6-aminohexyl)carbamate 182

tert-butyl (7-aminoheptyl)carbamate 183

tert-butyl (8-aminooctanyl)carbamate 184

tert-butyl (2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl) carbamate

Compound 181

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=7.3 Hz, 2H), 7.49 (d, J=7.1 Hz,2H), 6.89 (d, J=6.9 Hz, 2H), 6.70 (d, J=8.8 Hz, 1H), 6.48 (s, 1H), 5.72(s, 1H), 4.67 (s, 1H), 4.36 (s, 2H), 4.08-3.72 (m, 5H), 3.70-3.28 (m,5H), 3.27-2.87 (m, 5H), 2.78 (s, 1H), 2.37 (s, 3H), 1.51-0.91 (m, 11H).MS (ESI) m/z: 934.1 (M+H⁺).

Compound 182

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.5 Hz, 2H), 7.50 (t, J=9.2 Hz,2H), 6.90 (d, J=9.1 Hz, 2H), 6.70 (dd, J=9.0, 2.5 Hz, 1H), 6.57 (s, 1H),5.69 (d, J=26.8 Hz, 2H), 4.67 (d, J=6.1 Hz, 1H), 4.43-4.26 (m, 2H),4.03-3.75 (m, 5H), 3.70-3.33 (m, 5H), 3.32-3.10 (m, 4H), 3.03 (dd,J=12.9, 6.4 Hz, 2H), 2.75 (s, 1H), 2.37 (s, 3H), 1.32 (dd, J=41.6, 34.4Hz, 13H). MS (ESI) 948.1 (M+H⁺).

Compound 183

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.4 Hz, 2H), 7.50 (t, J=8.6 Hz,2H), 7.00-6.83 (m, 2H), 6.70 (dd, J=9.0, 2.4 Hz, 1H), 6.58 (s, 1H), 5.73(s, 1H), 5.62 (s, 1H), 4.68 (d, J=7.1 Hz, 1H), 4.37 (qd, J=10.7, 3.5 Hz,2H), 3.99-3.76 (m, 5H), 3.63 (s, 2H), 3.59-3.32 (m, 3H), 3.30-2.86 (m,6H), 2.84-2.65 (m, 1H), 2.37 (s, 3H), 1.55-0.93 (m, 15H). MS (ESI) m/z:962.1 (M+H⁺).

Compound 185

Compound 185 was prepared by a procedure similar to the one described inExample 31 by replacing ethanol in the last step with isopropanol(Propan-2-ol).

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=7.8 Hz, 2H), 7.49 (d, J=8.0 Hz,2H), 6.89 (d, J=7.1 Hz, 2H), 6.70 (d, J=8.3 Hz, 1H), 6.58 (s, 1H), 5.73(s, 2H), 5.28 (d, J=57.0 Hz, 2H), 4.68 (s, 1H), 4.01-3.75 (m, 5H),3.74-3.34 (m, 5H), 3.32-2.93 (m, 5H), 2.74 (s, 1H), 2.38 (s, 3H), 2.02(s, 1H), 1.48-1.13 (m, 15H). MS (ESI) 474.7[(M+2H⁺)/2]

Example 32 Synthesis of Compound 190

186 is coupled with indomethacin to give the amide 187 as described inexample 5. Compound 187 is dissolved in methanol and hydrogenated overPd/C to give compound 188. Compound 188 is then coupled with acid 173and esterified to give compound 190 as described in example 31.

Example 33 Synthesis of Compound 193

Compound 176 is synthesized as described in Example 31. 176 is coupledwith S, S -ethylenedicysteine 172 (CAS#14344-48-0) according toliterature procedures, see e.g. Yang et al., US 2006/6692724. Theethylester 192 is obtained by treating 191 with EtOH/TMSCl or EtOH/HCl.Compound 193 is prepared from compound 192 by treating with sodiumpertechnetate as described in Example 3.

The following compounds 194-201 can also be prepared according to theprocedure given in Example 33 by replacing compound 176 with theappropriate intermediate (synthesis described in examples 31-32.

Ex- Y = ample 194

195

196

197

198

199

200

201

Example 34 Synthesis of Compound 204

Compound 204 was prepared by the following procedure:

A solution of tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate 174(50 mg, 0.2 mmol), Re(CO)₅Br (82 mg, 0.2 mmol), and picolinaldehyde (22mg, 0.2 mmol) in methanol (20 ml) was stirred at 80° C. for 12 h undernitrogen. The reaction mixture was then concentrated and purified bycolumn chromatography to give compound 202 (110 mg, 79%). A solution ofHCl in dioxane (4N, 10 ml) and compound 202 (110 mg, 0.16 mmol) wasstirred at rt for 2 h, and then concentrated in vacuo to give crudeproduct 203 (100 mg), which was used directly in the next step.

To a solution of indomethacin (57 mg, 0.16 mmol) in DCM (20 ml) wasadded HOBt (22 mg, 0.16 mmol), EDCI (31 mg, 0.16 mmol), and DIEA (62 mg,0.48 mmol). The mixture was stirred at RT for 0.5 h, after which 203(100 mg, 0.16 mmol) was added. The reaction mixture was stirred at RTovernight. The mixture was then diluted with water (30 ml) and extractedwith EtOAc (25 ml×3). The combined organic layers were washed with brine(15 ml×2), dried over Na₂SO₄, filtered and the filtrate was concentratedand purified by prep-HPLC (Column: Acquity BEH C18, Waters Corp.;solvent A: water, solvent B: MeCN) to give the desired product 204.

¹H NMR (400 MHz, CDCl3) δ 9.00 (d, J=5.3 Hz, 1H), 8.66 (s, 1H),8.09-7.97 (m, 1H), 7.88 (d, J=7.6 Hz, 1H), 7.67 (d, J=8.5 Hz, 2H),7.60-7.44 (m, 3H), 6.98-6.84 (m, 2H), 6.69 (dd, J=9.0, 2.5 Hz, 1H), 6.00(s, 1H), 4.32-4.11 (m, 2H), 4.04-3.89 (m, 2H), 3.86-3.76 (m, 3H), 3.61(s, 2H), 3.57-3.38 (m, 6H), 3.38-3.22 (m, 2H), 2.39 (d, J=7.7 Hz, 3H).MS (ESI) m/z: 927.2 (M+H⁺).

The following compounds 205-211 can also be prepared according to theprocedure given in Example 34 by replacing tert-butyl2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate 174 with the appropriatereagent given in Table 2.

TABLE 2 Compound Y = Reagent used 205

tert-butyl (2-aminoethyl)carbamate 206

tert-butyl (2-(2- aminoethoxy)ethyl)carbamate 207

tert-butyl (6-aminohexyl)carbamate 208

tert-butyl (7-aminoheptyl)carbamate 209

tert-butyl (8-aminooctanyl)carbamate 210

tert-butyl (2-(2-(2-(2- aminoethoxy)ethoxy)ethoxy) ethyl)carbamate 211

tert-butyl (4- (aminomethyl)benzyl)carbamate

Compound 207

¹H NMR (400 MHz, DMSO-d₆) δ 9.19 (s, 1H), 9.03 (d, J=5.0 Hz, 1H),8.43-8.20 (m, 2H), 8.06 (s, 1H), 7.86-7.73 (m, 1H), 7.66 (dd, J=18.1,8.4 Hz, 4H), 7.13 (s, 1H), 6.93 (d, J=8.9 Hz, 1H), 6.70 (d, J=9.0 Hz,1H), 4.29-4.07 (m, 1H), 4.06-3.92 (m, J=6.5 Hz, 2H), 3.75 (s, 3H), 3.48(s, 2H), 3.17 (d, J=5.2 Hz, 2H), 2.22 (s, 3H), 1.90 (s, 2H), 1.52-1.14(m, J=36.9 Hz, 6H). MS (ESI) m/z: 851.2 (M+H⁺).

Compound 208

¹H NMR (400 MHz, CDCl3) δ 9.04 (d, J=5.1 Hz, 1H), 8.71 (s, 1H), 8.05(dd, J=7.7, 6.5 Hz, 1H), 7.92 (d, J=7.6 Hz, 1H), 7.67 (d, J=8.5 Hz, 2H),7.58-7.53 (m, 1H), 7.50 (t, J=8.0 Hz, 2H), 6.87 (t, J=5.3 Hz, 2H), 6.69(dd, J=9.0, 2.5 Hz, 1H), 5.59 (s, 1H), 4.37-4.14 (m, 1H), 4.07-3.97 (m,1H), 3.94-3.71 (m, 3H), 3.62 (s, 2H), 3.20 (dd, J=13.0, 6.7 Hz, 2H),2.37 (s, 3H), 2.15-1.92 (m, 2H), 1.52-1.14 (m, 8H). MS (ESI) m/z: 910.2(M+H⁺).

Compound 209

¹H NMR (400 MHz, CDCl3) δ 9.04 (d, J=5.1 Hz, 1H), 8.70 (s, 1H),8.12-8.00 (m, 1H), 7.90 (d, J=7.7 Hz, 1H), 7.67 (d, J=8.4 Hz, 2H),7.61-7.53 (m, 1H), 7.49 (d, J=8.4 Hz, 2H), 6.87 (t, J=5.7 Hz, 2H), 6.70(dd, J=9.0, 2.5 Hz, 1H), 5.59 (t, J=5.8 Hz, 1H), 4.34-4.12 (m, 1H), 4.02(dt, J=11.8, 7.4 Hz, 1H), 3.82 (s, 3H), 3.62 (s, 2H), 3.34-3.13 (m, 2H),2.38 (s, 3H), 2.23-1.85 (m, 2H), 1.47-1.11 (m, 10H). MS (ESI) m/z: 923.2(M+H⁺).

Compound 210

¹H NMR (400 MHz, CDCl3) δ 9.08-8.96 (m, 1H), 8.75 (s, 1H), 8.01 (td,J=7.8, 1.4 Hz, 1H), 7.88 (d, J=7.7 Hz, 1H), 7.67 (d, J=8.5 Hz, 2H), 7.54(dd, J=9.5, 3.8 Hz, 1H), 7.49 (d, J=8.5 Hz, 2H), 7.01-6.81 (m, 2H), 6.68(dd, J=9.0, 2.5 Hz, 1H), 6.07 (s, 1H), 4.25 (ddd, J=16.8, 16.2, 8.2 Hz,2H), 4.03 (dt, J=8.7, 6.3 Hz, 2H), 3.83 (d, J=11.1 Hz, 3H), 3.70-3.59(m, 3H), 3.55-3.34 (m, 8H). MS (ESI) 971.2 (M+H⁺).

Compound 211

¹H NMR (400 MHz, DMSO-d₆) δ 9.37 (s, 1H), 9.01 (d, J=5.2 Hz, 1H), 8.55(t, J=5.7 Hz, 1H), 8.31 (d, J=3.2 Hz, 2H), 7.77 (dd, J=8.8, 5.2 Hz, 1H),7.66 (dd, J=24.0, 8.4 Hz, 4H), 7.36 (d, J=7.9 Hz, 2H), 7.27 (d, J=7.9Hz, 2H), 7.14 (d, J=2.0 Hz, 1H), 6.96 (d, J=9.0 Hz, 1H), 6.71 (dd,J=8.9, 2.1 Hz, 1H), 5.76 (s, 2H), 5.37 (d, J=13.6 Hz, 1H), 5.09 (d,J=13.3 Hz, 1H), 4.31 (d, J=5.6 Hz, 2H), 3.73 (s, 3H), 3.59 (s, 2H), 2.22(s, 3H). MS (ESI) 915.2 (M+H⁺).

The following compounds 212-219 can also be prepared according to theprocedure given in Example 34 and for compounds 204-211, by substitutingRe(CO)₅Br (CAS No. 14220-21-4) in step 1 of example 34 with Re(CO)₅Cl(CAS No. 14099-01-5).

Com- pound Y = 212

213

214

215

216

217

218

219

Compound 219

¹H NMR (400 MHz, CDCl3) δ 8.98 (d, J=5.2 Hz, 1H), 8.69 (s, 1H), 8.03 (t,J=7.7 Hz, 1H), 7.87 (d, J=7.7 Hz, 1H), 7.68 (d, J=8.5 Hz, 2H), 7.63-7.52(m, 1H), 7.49 (d, J=8.4 Hz, 2H), 6.89 (dd, J=11.2, 5.7 Hz, 2H), 6.69(dd, J=9.0, 2.4 Hz, 1H), 6.05 (s, 1H), 4.33-4.15 (m, 2H), 3.95 (qd,J=10.8, 4.3 Hz, 2H), 3.81 (s, 3H), 3.61 (s, 2H), 3.58-3.23 (m, 8H), 2.38(s, 3H). MS (ESI) 883.2 (M+H⁺).

Example 35 Synthesis of Compound 221

Compound 176 is synthesized as described in Example 31. 176 is coupledwith picolinaldehyde to give the imine 220 according to literatureprocedures or methods well known to a person skilled in the art. Forexample, dissolving equal amounts in toluene and heating to reflux willaccomplish such a transformation. Compound 221 is prepared from compound220 as described by e.g. Alberto, R., Schibli, R., Schubiger, A. P., J.Am. Chem. Soc., 121, 6076-6077, (1999).

The following compounds 222-229 can also be prepared according to theprocedure given in Example 35 by replacing compound 176 with theappropriate intermediate (synthesis described in examples 5, 6, 31-32.

Com- pound Y = 222

223

224

225

226

227

228

229

230

Example 36 Synthesis of Compound 234

The synthesis of compound 25 is described in Example 6.

Intermediate 231

To a solution of 25 (400 mg, 0.81 mmol) and picolinaldehyde (87 mg, 0.81mmol, 1 eq) in MeOH (10 ml) was added AcOH (0.3 ml), the mixture wasstirred at RT for 30 min, NaBH(OAC)₃ (687 mg, 3.24 mmol, 4eq) was added,the mixture was stirred at RT overnight. Water (20 ml) was added andextracted with ethyl acetate (30 ml×3), the combined organic layers werewashed with brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated to give the crude product which was purified by columnchromatography (DCM: MeOH=10:1) to get the product (120 mg, 27%) as ayellow solid. LC-MS: m/z=547.3 [M+1]⁺

Intermediate 232

To a solution of 231 (120 mg, 0.22 mmol) and TEA (45 mg, 0.44 mmol) inTHF (10 mL) was added tert-butyl 2-bromoacetate (64 mg, 0.33 mmol). Themixture was stirred at 60° C. for 2 h. Water (10 ml) was added andextracted with ethyl acetate (3×30 ml), the combined organic layers werewashed with brine, dried over Na₂SO₄, concentrated to get the crudeproduct (130 mg, 90%) as yellow oil, which was used in the next stepwithout further purification. LC-MS: m/z=661.3 [M+1]⁺

Intermediate 233

To a solution of 232 (130 mg, 0.20 mmol) in DCM (10 ml) was added TFA (2ml), the mixture was stirred at RT for 4 h and then evaporated thesolvent to give the crude product (110 mg, 92%) as a yellow oil, whichwas used directly in the next step. LC-MS: m/z=605.3 [M+1]⁺.

233 (100 mg, 0.16 mmol) was dissolved in MeOH (5 ml), Re(CO)₅Cl (90 mg,0.25 mmol) was added and the mixture was stirred at 80° C. overnight.The solvent was evaporated and the residue was purified by Prep-HPLC(Column: Acquity BEH C18, Waters Corp, A: water/B: MeCN) to get theproduct 234 (20 mg, 14%) as a white solid. LC-MS: m/z=875.0[M+1]⁺. ¹HNMR (400 MHz, MeOD) δ 8.83 (d, J=5.2 Hz, 1H), 8.10 (td, J=7.8, 1.3 Hz,1H), 7.72 (d, J=8.5 Hz, 3H), 7.56 (dd, J=13.3, 7.5 Hz, 3H), 7.07 (d,J=2.4 Hz, 1H), 6.97 (d, J=9.0 Hz, 1H), 6.71 (dd, J=9.0, 2.5 Hz, 1H),4.67 (d, J=15.6 Hz, 1H), 4.51 (d, J=15.5 Hz, 1H), 3.89 (d, J=17.3 Hz,1H), 3.84 (s, 3H), 3.63 (s, 2H), 3.52 (ddd, J=31.5, 18.4, 11.2 Hz, 3H),3.26 (t, J=6.8 Hz, 2H), 2.35 (s, 3H), 1.91-1.67 (m, 2H), 1.65-1.53 (m,2H), 1.39 (d, J=3.1 Hz, 4H). ¹³C NMR (101 MHz, MeOD) δ 196.76, 195.90,181.92, 171.87, 168.59, 159.46, 156.23, 152.15, 140.25, 138.81, 135.75,134.32, 130.98, 130.82, 128.83, 125.57, 123.60, 114.62, 113.52, 111.33,101.17, 100.15, 69.68, 67.91, 60.58, 54.85, 38.88, 31.11, 28.89, 26.17,26.05, 24.79, 12.18.

Example 37 Synthesis of Compound 235

Compound 233 is prepared as described in Example 36.

233 is converted to 235 via the addition of [^(99m)Tc-(H₂O)₃(CO)₃]⁺(Isolink® kit; Mallinckrodt, St. Louis, Mo., USA) to a vial containing100m of compound 235 dissolved in 100 μl of water. The yield ofradiolabeled conjugate is maximized by adjusting the pH to 7 using 0.1 MHCl prior to heating at 80° C. for 1 hour. After incubation, themetallated complexes are purified by RP-HPLC and the peaks collectedinto either 100 μl of deionized H₂O containing 100m of bovine serumalbumin solution (BSA, in vitro analysis) or 100 μl of isotonic saline(in vivo analysis). Residual MeCN is evaporated from the solution usinga stream of nitrogen.

Example 38 Synthesis of Compound 237

Intermediate 236

To a solution of 25 (400 mg, 0.81 mmol) and picolinaldehyde (520 mg,4.86 mmol, 6 eq) in MeOH (20 ml) was added AcOH (0.3 ml), the mixturewas stirred at RT for 30 min. NaBH(OAc)₃ (687 mg, 3.24 mmol, 4eq) wasadded and the mixture was stirred at RT overnight. Water (20 ml) wasadded and the mixture was extracted with ethyl acetate (3×30 ml). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated to give the crude product,which was purified by column chromatography on silica gel(DCM:MeOH=10:1) to get the product (150 mg, 34%) as a yellow solid.LC-MS: m/z=638.0 [M+1]⁺.

To a solution of 236 (64 mg, 0.1 mmol) in MeOH (5 ml) was addedRe(CO)₅Cl (54.3 mg, 0.15 mmol, 1.5eq), the mixture was stirred at 80° C.overnight. The mixture was then concentrated and purified by Prep-HPLC(Column: Acquity BEH C18, Waters Corp, A: water/B: MeCN) to give theproduct 237 (20 mg, 23%) as a white solid. LC-MS: m/z=908.1 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.77-8.76 (d, J=5.5 Hz, 2H), 7.84-7.81 (m, 2H),7.60 (d, J=8.6 Hz, 2H), 7.48-7.42 (dd, J=20.2, 8.2 Hz, 4H), 7.32-7.21(m, 2H), 6.96 (d, J=2.4 Hz, 1H), 6.86 (d, J=9.0 Hz, 1H), 6.60-6.57 (dd,J=9.0, 2.5 Hz, 1H), 4.78-4.63 (m, 4H), 3.73 (s, 3H), 3.69-3.59 (m, 2H),3.52 (s, 2H), 3.16-3.14 (t, J=6.9 Hz, 2H), 2.23 (s, 3H), 1.76 (s, 2H),1.56-1.42 (m, 2H), 1.32 (s, 4H).

Example 39 Synthesis of Compound 238

Compound 236 is prepared as described in example 38.

236 is converted to 238 by a procedure similar to the one described inexample 37.

Example 40 Synthesis of Compound 242

Cyclopentadienyltricarbonylrhenium(I) carboxylic acid 241 is synthesizedas described by Siden Top, Jean-Sébastien Lehn, Pierre Morel, GerardJaouen, J. Organomet. Chem., 583, 63-68, (1999). To a solution ofcompound 25 (example 6) (0.16 mmol) in DCM (20 ml) was added HOBt (22mg, 0.16 mmol), EDCI (31 mg, 0.16 mmol), and DIEA (62 mg, 0.48 mmol).The mixture was stirred at RT for 0.5 h, after which 241 (0.16 mmol) wasadded. The reaction mixture was stirred at RT overnight. The mixture wasthen diluted with water (30 ml) and extracted with EtOAc (25 ml×3). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated and purified by prep-HPLC(Column: Acquity BEH C18, Waters Corp, A: water, B: MeCN) to give thedesired product 242.

¹H NMR (400 MHz, MeOD) δ 8.03 (s, 1H), 7.72 (d, J=8.5 Hz, 2H), 7.58 (d,J=8.5 Hz, 2H), 7.04 (d, J=2.4 Hz, 1H), 6.95 (d, J=9.0 Hz, 1H), 6.69 (dd,J=9.0, 2.4 Hz, 1H), 6.17 (t, J=4.0 Hz, 2H), 5.57 (t, J=4.0 Hz, 2H), 3.82(s, 3H), 3.63 (s, 2H), 3.24-3.18 (m, 4H), 2.34 (s, 3H), 1.61-1.44 (m,4H), 1.41-1.27 (m, 4H). MS (ESI) m/z: 818.0[M+H⁺].

The following compounds 243-251 can also be prepared according to theprocedure given in Example 40 by replacing compound 25 with theappropriate intermediate (synthesis described in examples 5, 31, and32).

Com- pound Y = 243

244

245

246

247

248

249

250

251

Compound 243

¹H NMR (400 MHz, DMSO-d₆) δ 8.31 (t, J=5.2 Hz, 1H), 8.07 (t, J=5.2 Hz,1H), 7.67 (dd, J=22.2, 8.6 Hz, 4H), 7.09 (d, J=2.4 Hz, 1H), 6.94 (d,J=9.0 Hz, 1H), 6.70 (dd, J=9.0, 2.5 Hz, 1H), 6.20 (t, J=2.2 Hz, 2H),5.76-5.68 (m, 2H), 3.76 (s, 3H), 3.50 (s, 2H), 3.23-3.09 (m, 4H), 2.22(s, 3H). MS (ESI) m/z: 762.0 [M+H⁺].

Compound 244

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.5 Hz, 2H), 7.50 (d, J=6.9 Hz,2H), 6.91 (d, J=2.3 Hz, 1H), 6.88 (d, J=9.0 Hz, 1H), 6.71 (dd, J=9.0,2.3 Hz, 2H), 6.12 (t, J=2.0 Hz, 2H), 5.95 (s, 1H), 5.34 (t, J=2.0 Hz,2H), 3.83 (s, 3H), 3.68 (s, 2H), 3.53-3.35 (m, 8H), 2.39 (s, 3H). MS(ESI) 806.0[M+H⁺].

Compound 245

¹H NMR (400 MHz, CDCl3) δ 7.63 (d, J=8.2 Hz, 3H), 7.50 (d, J=8.3 Hz,2H), 7.17 (d, J=8.2 Hz, 2H), 7.04 (d, J=8.9 Hz, 1H), 6.84-6.73 (m, 4H),6.12 (s, 2H), 5.45 (s, 2H), 5.32 (s, 1H), 3.83 (s, 3H), 3.55 (s, 2H),3.44 (s, 2H), 2.66 (s, 2H), 2.14 (s, 3H). MS (ESI) m/z: 860.0 [M+H⁺].

Compound 246

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4 Hz,2H), 6.87 (d, J=9.3 Hz, 2H), 6.71 (dd, J=9.0, 2.2 Hz, 1H), 6.01 (s, 1H),5.96 (s, 2H), 5.61 (s, 1H), 5.34 (s, 2H), 3.82 (s, 3H), 3.64 (s, 2H),3.31 (d, J=5.7 Hz, 2H), 3.21 (d, J=5.9 Hz, 2H), 2.39 (s, 3H), 1.42 (m,4H), 1.22 (m, 6H). MS (ESI) m/z: 832.2 [M+H⁺].

Compound 247

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=7.9 Hz, 2H), 7.49 (d, J=7.7 Hz,2H), 6.87 (d, J=9.1 Hz, 2H), 6.70 (d, J=8.0 Hz, 1H), 5.90 (s, 2H), 5.61(s, 1H), 5.32 (d, J=16.8 Hz, 3H), 3.82 (s, 3H), 3.64 (s, 2H), 3.25 (dd,J=41.8, 5.8 Hz, 4H), 2.39 (s, 3H), 1.59-1.23 (m, 8H). MS (ESI)846.2[M+H⁺].

Compound 248

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.5 Hz, 2H), 7.50 (t, J=7.8 Hz,2H), 6.93-6.86 (m, 2H), 6.70 (dd, J=9.0, 2.4 Hz, 1H), 6.61 (s, 1H), 6.08(s, 1H), 5.96 (s, 2H), 5.31 (t, J=4.0 Hz, 2H), 3.82 (s, 3H), 3.65 (s,2H), 3.59-3.37 (m, 16H), 2.38 (s, 3H). MS (ESI) m/z: 894.2 [M+H⁺].

Compound 249

¹H NMR (400 MHz, CDCl3) δ 7.64 (d, J=8.5 Hz, 2H), 7.47 (d, J=8.5 Hz,2H), 7.17 (d, J=8.0 Hz, 2H), 7.09 (d, J=8.0 Hz, 2H), 6.84 (dd, J=5.7,3.2 Hz, 2H), 6.68 (dd, J=9.1, 2.4 Hz, 1H), 6.09 (t, J=4.0 Hz, 1H),5.96-5.89 (m, 3H), 5.35 (t, J=4.0 Hz, 2H), 4.46 (d, J=5.6 Hz, 2H), 4.35(d, J=5.9 Hz, 2H), 3.78 (s, 3H), 3.68 (s, 2H), 2.36 (s, 3H). MS (ESI)m/z: 838.0 [M+H⁺].

Compound 250

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.3 Hz, 2H), 7.50 (d, J=8.3 Hz,2H), 7.04 (s, 1H), 6.87 (d, J=9.6 Hz, 2H), 6.72 (d, J=9.1 Hz, 1H), 6.10(s, 2H), 5.83 (s, 1H), 5.34 (s, 2H), 3.83 (s, 3H), 3.67 (s, 2H), 3.39(s, 2H), 3.26 (s, 2H), 2.39 (s, 3H), 1.51 (s, 3H). MS (ESI) m/z: 790.0[M+H⁺].

Compound 251

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.5 Hz, 2H), 7.50 (d, J=8.5 Hz,2H), 6.91 (d, J=2.3 Hz, 1H), 6.86 (d, J=9.0 Hz, 1H), 6.70 (dd, J=9.0,2.4 Hz, 1H), 6.60 (s, 1H), 6.00 (m, 3H), 5.33 (t, J=4.0 Hz, 2H), 3.82(s, 3H), 3.65 (s, 2H), 3.46 (m, 12H), 2.40 (s, 3H). MS (ESI) m/z: 850.1[M+H⁺].

Example 41 Synthesis of Compound 254

To a solution of compound 25 (example 6) (0.16 mmol) in DCM (20 ml) isadded HOBt (22 mg, 0.16 mmol), EDCI (31 mg, 0.16 mmol), and DIEA (62 mg,0.48 mmol). The mixture was stirred at RT for 0.5 h, after which 252(CAS No. 1271-42-7, Sigma-Aldrich, Saint Louis, Mo., USA; catalog no.106887; 0.16 mmol) is added. The reaction mixture is stirred at RTovernight. The mixture is then diluted with water and extracted withEtOAc (25 ml×3). The combined organic layers are washed with brine (15ml×2), dried over Na₂SO₄, filtered and the filtrate is concentrated andpurified by prep-HPLC (Column: Acquity BEH C18, Waters Corp, A: water,B: MeCN) to give the desired product 253.

253 can be converted into the Tc complex 254 as described in theliterature, see e.g. Bioorg. Med. Chem. Letters 22 (2012) 6352-6357; J.Med. Chem. (2007), 50, 543-549; J. Med. Chem. (2013), 56, 471-482; J.Med. Chem. (2014), 57, 7113-7125.

Example 42 Synthesis of Compound 260

Compound 255 is made according to the procedure in: Ziv, Ilan et al., WO2003/101948 and Mahmood, A., et al., WO 2001/083436. Compound 255 isalkylated with 1-bromo-6-chlorohexane to form 256 as previouslydescribed in: Mahmood, A.; et al., “Functionalized tetradentate chelatesand their Technetium and Rhenium complexes: synthesis spectroscopy andstructural characterization”, in Technetium and Rhenium in chemistry andnuclear medicine 5, pp. 253-257 (Nicolini, M. et al.) and also inMahmood, A., et al., WO 2001/083436. Compound 256 is treated withammonia in methanol at rt to give amine 257.

To a solution of indomethacin (57 mg, 0.16 mmol) in DCM (20 ml) wasadded HOBt (22 mg, 0.16 mmol), EDCI (31 mg, 0.16 mmol), and DIEA (62 mg,0.48 mmol). The mixture was stirred at RT for 0.5 h, after which (257)(85 mg, 0.16 mmol) was added. The reaction mixture was stirred at RTovernight. The mixture was then diluted with water (30 ml) and extractedwith EtOAc (25 ml×3). The combined organic layers were washed with brine(15 ml×2), dried over Na₂SO₄, filtered and the filtrate was concentratedto give the free dithiol 259.

Compound 259 is dissolved in DCM and treated with TFA in presence of5eq. of Et₃SiH at 80° C. for 4 h. The volatiles are removed bylyophilizing overnight. To a stirred solution of the residue (100 mg,0.14 mmol) in NMP (15 mL), (PPh₃)₂ReOCl₃ (Sigma-Aldrich, Order #370193,106 mg, 0.127 mmol) was added. The mixture was stirred at 80° C. for 16h. The solution was purified by Prep-HPLC (Column: Acquity BEH C18,Waters Corp, solvent A: Water, NH₄HCO₃, solvent B: MeCN) to givecompound 260 as a solid.

1H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.5 Hz, 2H), 7.50 (d, J=8.5 Hz,2H), 6.91-6.83 (m, 2H), 6.71 (d, J=9.0, 2.4 Hz, 1H), 5.60 (s, 1H),4.65-4.53 (m, 2H), 4.15-4.03 (m, 2H), 3.94-3.86 (m, 1H), 3.83 (s, 3H),3.65 (s, 2H), 3.49-3.39 (m, 1H), 3.40-3.11 (m, 6H), 2.85 (dd, J=13.4,4.2 Hz, 1H), 2.40 (s, 3H), 1.62-1.51 (m, 2H), 1.48-1.38 (m, 2H),1.37-1.19 (m, 4H). MS (ESI) m/z: 855.0 (M+Na⁺)

The following compounds 261-269 can also be prepared according to theprocedure given in Example 42 by replacing 1-bromo-6-chloro-hexane inExample 42 with the appropriate dihalide.

Compound Y = 261

262

263

264

265

266

267

268

269

Compound 261

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.3 Hz,2H), 6.87 (d, J=9.5 Hz, 1H), 6.78-6.64 (m, 1H), 5.77 (s, 1H), 4.70-4.42(m, 2H), 4.07 (dd, J=10.8, 4.5 Hz, 1H), 3.97 (d, J=16.5 Hz, 1H), 3.83(s, 3H), 3.67 (s, 2H), 3.56-3.42 (m, 1H), 3.38-2.95 (m, 6H), 2.81 (dd,J=13.6, 4.1 Hz, 1H), 2.40 (s, 3H), 1.85-1.40 (m, 5H). MS (ESI) m/z:805.0 (M+H⁺)

Compound 262

¹H NMR (400 MHz, CDCl3) δ 7.69 (d, J=8.2 Hz, 2H), 7.50 (d, J=8.3 Hz,2H), 6.92-6.85 (m, 2H), 6.72 (s, 1H), 6.02 (s, 1H), 4.57-4.55 (m, 1H),4.46-4.41 (m, 1H), 4.11-4.08 (m, 1H), 3.84-3.80 (m, 5H), 3.68-3.52 (m,5H), 3.51-3.41 (m, 4H), 3.41-3.09 (m, 4H), 2.66 (d, J=12.8 Hz, 1H), 2.42(d, J=4.1 Hz, 3H). MS (ESI) m/z: 843.0 (M+Na⁺).

Compound 263

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.3 Hz, 2H), 7.50 (d, J=8.4 Hz,2H), 6.90-6.78 (m, 2H), 6.72 (d, J=9.1 Hz, 1H), 5.73 (s, 1H), 4.70-4.49(m, 2H), 4.19-4.00 (m, 2H), 3.83 (s, 3H), 3.78 (s, 1H), 3.69 (s, 2H),3.45-3.05 (m, 7H), 2.85 (dd, J=13.2, 4.1 Hz, 1H), 2.41 (s, 3H),1.87-1.66 (m, 2H), 1.62-1.45 (m, 2H), 1.31-1.15 (m, 2H). MS (ESI) m/z:840.9 (M+Na⁺).

Compound 264

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.2 Hz, 2H), 7.50 (d, J=7.8 Hz,2H), 6.96-6.79 (m, 2H), 6.71 (d, J=8.8 Hz, 2H), 5.58 (s, 1H), 4.70-4.51(m, 2H), 4.20-4.01 (m, 2H), 3.98-3.88 (m, 1H), 3.82 (s, 1H), 3.64 (s,1H), 3.52-3.41 (m, 1H), 3.40-3.04 (m, 6H), 2.85 (d, J=9.7 Hz, 1H), 2.40(s, 3H), 1.85-1.67 (m, 2H), 1.47-1.37 (m, 2H), 1.37-1.12 (m, 6H). MS(ESI) m/z: 847.1 (M+H⁺).

Compound 265

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.5 Hz, 2H), 7.49 (d, J=8.5 Hz,2H), 6.87 (dd, J=8.8, 5.7 Hz, 2H), 6.70 (dd, J=9.0, 2.4 Hz, 1H), 5.58(t, J=5.7 Hz, 1H), 4.71-4.51 (m, 2H), 4.16-4.02 (m, 2H), 4.00-3.88 (m,1H), 3.82 (s, 3H), 3.64 (s, 2H), 3.55-3.42 (m, 1H), 3.41-3.30 (m, 1H),3.29-3.08 (m, 5H), 2.85 (dd, J=13.4, 4.3 Hz, 1H), 2.39 (s, 3H),1.85-1.66 (m, 2H), 1.64-1.56 (m, 2H), 1.46-1.35 (m, 2H), 1.36-1.12 (m,8H). MS (ESI) m/z: 883.4 (M+Na⁺).

Compound 266

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.2 Hz, 2H), 7.54-7.44 (m, 2H),6.99-6.85 (m, 2H), 6.74 (d, J=9.3 Hz, 1H), 6.11 (s, 1H), 4.70-4.49 (m,2H), 4.22-4.01 (m, 4H), 3.84 (d, J=3.3 Hz, 4H), 3.71 (s, 2H), 3.59-3.12(m, 10H), 2.87 (d, J=13.5 Hz, 1H), 2.35 (s, 3H), 1.91 (s, 2H), 1.66-1.54(m, 2H). MS (ESI) m/z: 849.1 (M+H⁺).

Compound 267

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.5 Hz, 2H), 7.50 (d, J=7.8 Hz,2H), 6.92-6.81 (m, 2H), 6.71 (dd, J=9.0, 2.5 Hz, 1H), 5.88 (s, 1H), 4.75(d, J=16.9 Hz, 1H), 4.63-4.51 (m, 1H), 4.20 (d, J=16.8 Hz, 1H),4.14-4.05 (m, 1H), 4.04-3.94 (m, 1H), 3.84 (s, J=16.4 Hz, 3H), 3.78 (s,1H), 3.72-3.57 (m, 5H), 3.55-3.06 (m, 8H), 2.97-2.69 (m, 1H), 2.40 (s,3H), 1.86-1.68 (m, 2H). MS (ESI) m/z: 835.1 (M+H⁺).

Compound 268

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=7.5 Hz, 2H), 7.50 (d, J=7.6 Hz,2H), 7.03-6.93 (m, 1H), 6.90 (s, 1H), 6.73 (d, J=8.6 Hz, 1H), 4.72 (d,J=11.8 Hz, 1H), 4.59 (d, J=6.4 Hz, 1H), 4.22-3.94 (m, 3H), 3.83 (s, 3H),3.66 (s, 3H), 3.45 (d, J=38.6 Hz, 3H), 3.33-3.06 (m, 3H), 2.88 (s, 3H),2.37 (s, 5H), 2.01 (s, 2H), 1.66 (s, 2H). MS (ESI) m/z: 897.1 (M+H⁺).

Compound 269

¹H NMR (400 MHz, CDCl3) δ 7.65 (d, J=8.4 Hz, 2H), 7.47 (d, J=8.4 Hz,2H), 6.95 (s, 2H), 6.89 (d, J=9.0 Hz, 1H), 6.69 (dd, J=8.9, 2.1 Hz, 1H),4.68 (d, J=17.4 Hz, 1H), 4.43-4.32 (m, 1H), 4.17 (d, J=17.4 Hz, 1H),4.09-3.98 (m, 1H), 3.95-3.76 (m, 6H), 3.76-3.70 (m, 1H), 3.66 (d, J=6.5Hz, 2H), 3.62-3.41 (m, 7H), 3.36-3.21 (m, 3H), 3.17-3.05 (m, 2H), 2.80(dd, J=12.8, 4.0 Hz, 1H), 2.37 (s, 1H). MS (ESI) m/z: 886.9 (M+Na⁺).

Example 43 Synthesis of Compound 270

Compound 270 is synthesized by reacting 259 (example 42) with^(99m)Tc-glucoheptonate as described in: Mahmood, A., et al., WO2001/083436.

Example 44 Synthesis of Compound 275

Compound 255 is made according to the procedure in: Ziv, Ban et al., WO2003/101948 and Mahmood, A., et al., WO 2001/083436. 255 is reduced withLiAlH₄ and Boc-protected to give compound 271 according to the proceduredescribed in Ono, M., et al., ACS Chem. Neurosci., 1, 598-607, (2010).

Intermediate 272

To a solution of indomethacin(2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3-yl)acetic acid,1.5 g, 4.2 mmol) in DMF (20 mL) was added 6-aminohexan-1-ol (589 mg,5.04 mmol), HOBt (1.02 g, 7.56 mmol), EDCI (1.45 g, 7.56 mmol) and DIPEA(2.16 g, 16.8 mmol). The resulting solution was stirred RT overnight bywhich time LCMS showed completion of the reaction. Then water was added(80 mL), and the mixture was extracted with EtOAc (50 mL×3). Thecombined organic layers were washed with brine, dried over Na₂SO₄filtered and the filtrate was concentrated to obtain the product (1.5 g,yield: 78%) as a light yellow solid.

Intermediate 273

To a solution of 272 (1.5 g, 3.28 mmol) in DCM (30 ml) was addedDess-Martin periodinane(1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one, 2.09 g,4.93 mmol) at 0° C. The resulting solution was stirred RT for 2 h, thena sat'd. aq. solution of Na₂S₂O₃ (10 mL), was added. The mixture wasextracted with DCM (20 mL×3). The combined organic layers were washedwith brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated to get the crude product 273 (1.2 g, yield: 81%) as ayellow solid which was used in the next step without furtherpurification.

Intermediate 274

To a solution of 273 (1.2 g, 2.64 mmol) in MeOH (30 mL) was addedcompound 271 (1.0 g, 1.98 mmol), and 1 mL of CH₃COOH. The resultingsolution was stirred at rt for 0.5 h. Then, NaBH₃CN (332 mg, 5.28 mmol)was added and the mixture was stirred for another 2 h. Then the reactionwas quenched with water (20 mL) and extracted with DCM (30 mL×4). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated to give the crude product,which was purified by chromatography on silica gel (DCM:MeOH=50:1) toget the product 274 (1.2 g yield: 63%) as a yellow oil.

Compound 275

To a solution of 274 (1.2 g, 1.26 mmol) in TFA (20 mL) was added Et3SiH(220 mg, 1.9 mmol). The resulting mixture was stirred at 80° C. for 2 h.Then volatiles were removed and the residue was dissolved in NMP (10mL). (PPh₃)₂ReOCl₃ (Sigma-Aldrich, Order #370193, 1.0 eq) was added andthe mixture was stirred at 80° C. overnight. The reaction wasconcentrated and the residue was purified by Prep-HPLC (Column: AcquityBEH C18, Waters Corp, A: water/B: MeCN) to give 275 as a light pinksolid (80 mg, 8% yield over both steps).

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4 Hz,2H), 6.98-6.80 (m, 2H), 6.70 (dd, J=9.0, 2.3 Hz, 1H), 5.61 (s, 1H), 4.12(dt, J=12.7, 6.3 Hz, 2H), 3.99 (dd, J=18.6, 7.5 Hz, 2H), 3.91-3.73 (m,5H), 3.64 (s, 2H), 3.51-3.16 (m, 7H), 3.11-2.88 (m, 2H), 2.82-2.61 (m,1H), 2.40 (s, 3H), 1.69 (dd, J=11.9, 7.1 Hz, 2H), 1.51-1.36 (m, 2H),1.29 (s, 4H). MS (ESI) m/z: 818.9 [M+H⁺].

Example 45 Synthesis of Compound 276

274 is converted into the dithiol by reacting with TFA and Et₃SiH asdescribed for compound 275. The ^(99m)Tc complex 276 is prepared by aligand exchange reaction employing ^(99m)Tc-glucoheptonate as describedin e.g.: Ono, M., et al., ACS Chem. Neurosci., 1, 598, (2010) and Ono,M., et al., Bioorg. Med. Chem. Lett., 20, 5743-5748, (2010).

Example 46 Synthesis of Compound 283

Compound 277 can be transformed to 278 by reacting 277 withdi-tert-butyldicarbonate in the presence of a base such as NaHCO₃ or aq.NaOH. The nitrile 279 is obtained using a procedure similar to the onedescribed in: Mizuno, A.; Hamada, Y.; Shioiri, T., Synthesis 1007,(1980). The nitrile is reduced with hydrogen in presence of a catalyst,such as Raney®-Nickel (W.R. Grace and Co.) in a solvent such as methanolor tert-butanol to give amine 280. Indomethacin is coupled to compound280 as described in example 5 and treated with TFA to give the amide281. 281 is coupled with the S-benzoyl-protected thioglycolic acid usingDCC as the coupling agent to afford ligand 282. 283 is made followingthe procedure in Ono, et al. Bioorg. Med. Chem. Lett. (2010), 20,5743-5748.

Example 47 Synthesis of Compound 284

The Technetium complex 284 is prepared from compound 282 (example 46) asdescribed in: Eisenhut, M., et al., J. Med. Chem., 45, 5802-5805,(2002).

Example 48 Synthesis of Compound 286

Compound 282 (Example 46) is reduced with LiAlH₄ in a solvent such asTHF to give compound 285. 286 is prepared from 285 following theprocedure in Ono, et al. Bioorg. Med. Chem. Lett. (2010), 20, 5743-5748.

Example 49 Synthesis of Compound 287

The Technetium complex 287 is prepared from compound 285 (example 48) asdescribed in: Eisenhut, M., et al., J. Med. Chem., 45, 5802-5805,(2002).

Example 50 Synthesis of compounds 290 and 291

Compound 281 (Example 46) is coupled with tert-butyl bromoacetate inpresence of a base, such as NaH, to afford compounds 288 and 289, whichare separated by column chromatography on silica gel. 288 is transformedto 290 by treating with TFA in DCM followed by reaction with Re(CO)₅Clby a procedure similar to the one described in Example 36. Similarly,291 can be obtained form 289.

Example 51 Synthesis of compounds 292 and 293

288 and 289 are converted to 292 and 293, respectively, by a proceduresimilar to the one described in example 37.

Example 52 Synthesis of Compound 297

Intermediate 294

To the mixture of 18 (1.07 g, 3 mmol) and NH₄Cl (318 mg, 6 mmol) in DMF(40 ml) was added HOBt (607 mg, 4.5 mmol), EDCI (864 mg, 4.5 mmol) andDIPEA (1.6 ml, 9 mmol). The resulting mixture was stirred at roomtemperature overnight. Water (100 ml) was added and the product wasextracted with EtOAc (2×100 ml). The combined organic layers were driedover Na₂SO₄, and the solvent was removed under reduced pressure to givethe product without further purification as a yellow oil (1 g, 93.6%).LC-MS: m/z=357.1 [M+H]

Intermediate 295

To the solution of compound 294 (1 g, 2.8 mmol) in DCM (20 ml) was addedTEA (606 mg, 6 mmol) and TFAA (2.1 g, 10 mmol) at 0° C. The resultingsolution was stirred at RT for 1 h. The reaction solution wasconcentrated under reduced pressure to give a yellow solid. The crudesolid was washed with hexane (50 ml) to give the product 295 as a yellowsolid (700 mg, 74%). LC-MS: m/z=339.2 [M+H]

Intermediate 296

The mixture of compound 295 (700 mg, 2.07 mmol) and Raney®-Nickel (W.R.Grace and Co., 100 mg) in THF/MeOH (5 ml/20 ml) was stirred under ahydrogen atmosphere overnight. The catalyst was removed by filteringthrough a column of Celite® (J. T. Baker, Phillipsberg, N.J.,diatomaceous earth) and the filtrate was concentrated under reducedpressure to give the crude product, which was purified by prep-HPLC(Column: Acquity BEH C18, Waters Corp, solvent A: water/solvent B: MeCN)to give the product as light yellow solid (500 mg, 70%). LC-MS:m/z=343.1 [M+H]⁺.

To a solution of compound 296 (171 mg, 0.5 mmol) and compound 241 (191mg, 0.5 mmol) in DMF (15 ml) was added HOBt (135 mg, 1 mmol), EDCI (192mg, 1 mmol) and DIPEA (258 mg, 2 mmol). The resulting solution wasstirred at rt overnight. The reaction was quenched with saturated NaHCO₃solution (30 ml) and extracted with ethyl acetate (2×50 ml). Thecombined organic extracts were washed with brine (30 ml), dried overNa₂SO₄, concentrated, and purified by prep-HPLC (Column: Acquity BEHC18, Waters Corp, solvent A: water/solvent B: MeCN) to give the compound297 as a light yellow solid (20 mg, 6%). LC-MS: m/z=343.2 [M+H]⁺; ¹H NMR(400 MHz, DMSO-d6) δ 8.42 (t, J=5.8 Hz, 1H), 7.65 (q, J=8.6 Hz, 3H),7.10 (d, J=2.3 Hz, 1H), 7.01 (d, J=9.0 Hz, 1H), 6.72 (dd, J=9.0, 2.4 Hz,1H), 6.23 (t, J=2.1 Hz, 2H), 5.93-5.55 (m, 2H), 3.78 (s, 3H), 3.46-3.33(m, 2H), 2.82 (t, J=6.8 Hz, 2H), 2.15 (s, 3H).

Example 53 Synthesis of Compound 299

Compound 299 is prepared from compound 296 (example 52) by a proceduresimilar to the one described in Example 41.

Example 54 Synthesis of Compound 301

Compound 296 is transformed into the imine 300 by a procedure similar tothe one described for compound 220 in example 35. 300 is converted intothe Rhenium complex 301 as described in example 36.

Alternatively, compound 301 can be prepared as follows: A mixture of 296(100 mg, 0.29 mmol), picolinaldehyde (31 mg, 0.29 mmol) and Re(CO)₅Cl(105 mg, 0.29 mmol) in MeOH (10 mL) was stirred at 75° C. for 3 h, thencooled to rt. The precipitate was filtered and washed with MeOH (2×20ml), and dried to give compound 301 as a yellow solid (51 mg, 23%).LC-MS: m/z=759.9 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.32 (s, 1H), 9.06(d, J=5.2 Hz, 1H), 8.36 (t, J=7.7 Hz, 1H), 8.27 (d, J=7.5 Hz, 1H),7.92-7.76 (m, 1H), 7.68 (dd, J=28.7, 8.4 Hz, 4H), 7.23 (d, J=2.1 Hz,1H), 7.03 (d, J=9.0 Hz, 1H), 6.76 (dd, J=9.0, 2.2 Hz, 1H), 4.21 (t,J=7.9 Hz, 2H), 3.77 (s, 3H), 3.43-3.02 (m, 2H), 2.27 (s, 3H).

Example 55 Synthesis of Compound 302

Compound 300 (example 54) is converted to 302 by a procedure similar tothe one described in example 37.

The following compounds 303-307 can be prepared according to theprocedure given in Example 44 by replacing 6-aminohexan-1-ol with theappropriate aminoalcohol.

Compound Y = 303

304

305

306

307

Compound 303

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.2 Hz, 2H), 7.50 (d, J=8.3 Hz,2H), 7.01-6.83 (m, 2H), 6.77-6.63 (m, 1H), 5.78 (s, 1H), 4.22-4.00 (m,2H), 3.98-3.58 (m, 8H), 3.59-3.06 (m, 8H), 2.96 (td, J=11.7, 6.5 Hz,1H), 2.81 (d, J=11.4 Hz, 1H), 2.74-2.52 (m, 1H), 2.39 (s, 3H), 1.78-1.64(m, 2H), 1.46 (d, J=22.8 Hz, 2H). MS (ESI) m/z: 791.0 [M+H⁺].

Compound 304

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.3 Hz, 2H), 7.49 (d, J=8.2 Hz,2H), 6.99-6.77 (m, 2H), 6.71 (dd, J=9.0, 1.9 Hz, 1H), 5.71 (d, J=5.5 Hz,1H), 4.26-4.07 (m, 2H), 4.02-3.89 (m, 1H), 3.85-3.71 (m, 5H), 3.64 (s,2H), 3.51-3.13 (m, 8H), 3.09-2.87 (m, 2H), 2.71 (dd, J=13.1, 3.2 Hz,1H), 2.40 (s, 3H), 1.89-1.70 (m, 2H), 1.59-1.46 (m, 2H), 1.32-1.19 (m,2H). MS (ESI) m/z: 805.0 [M+H⁺].

Compound 305

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.5 Hz, 2H), 7.49 (d, J=8.5 Hz,2H), 6.99-6.78 (m, 2H), 6.70 (dd, J=9.1, 2.3 Hz, 1H), 5.59 (s, 1H),4.27-3.92 (m, 4H), 3.90-3.71 (m, 5H), 3.64 (s, 2H), 3.55-3.14 (m, 7H),3.06-2.88 (m, 2H), 2.79-2.65 (m, 1H), 2.39 (s, 3H), 1.81-1.68 (m, 2H),1.41 (dt, J=14.5, 7.3 Hz, 2H), 1.35-1.13 (m, 6H). MS (ESI) m/z: 833.2[M+H⁺].

Compound 306

1H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.4 Hz, 2H), 7.49 (d, J=8.4 Hz,2H), 7.10-6.79 (m, 2H), 6.71 (dd, 1H), 5.89 (s, 1H), 4.22-3.97 (m, 3H),3.97-3.74 (m, 5H), 3.63 (s, 2H), 3.58-3.07 (m, 11H), 3.05-2.94 (m, 1H),2.89 (d, J=11.6 Hz, 1H), 2.71 (d, J=9.5 Hz, 2H), 2.36 (s, 3H), 1.98-1.83(m, 2H), 1.78-1.66 (m, 2H). MS (ESI) m/z: 835.1 [M+H+].

Compound 307

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.3 Hz, 2H), 7.50 (d, J=8.3 Hz,2H), 7.05-6.78 (m, 2H), 6.71 (d, J=8.9 Hz, 1H), 5.60 (s, 1H), 4.36-3.94(m, 3H), 3.94-3.71 (m, 5H), 3.64 (s, 2H), 3.58-3.43 (m, 1H), 3.40-3.08(m, 6H), 3.09-2.89 (m, 2H), 2.84-2.65 (m, 1H), 2.40 (s, 3H), 2.13-1.88(m, 1H), 1.85-1.60 (m, 2H), 1.44-1.34 (m, 2H), 1.14-0.90 (m, 6H). MS(ESI) m/z: 833.1 [M+H⁺].

The following compounds 308-313 can also be prepared according to theprocedure given in Example 40 by replacing compound 25 with theappropriate intermediate (synthesis described in examples 5, 31, and32).

Compound Y= 308

309

310

311

312

313

Compound 308

¹H NMR (400 MHz, CDCl3) δ 7.66 (d, J=8.1 Hz, 2H), 7.49 (d, J=8.1 Hz,2H), 6.88 (d, J=8.8 Hz, 2H), 6.71 (d, J=8.8 Hz, 1H), 6.48 (s, 1H), 6.09(s, 2H), 5.81 (s, 1H), 5.33 (s, 2H), 3.82 (s, 3H), 3.65 (s, 2H),3.33-3.19 (m, 4H), 2.38 (s, 3H), 1.61-1.51 (m, 2H), 1.48-1.40 (m, 2H),1.36-1.23 (m, 2H). MS (ESI) m/z: 804.0 [M+H⁺].

Compound 309

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.1 Hz, 2H), 7.49 (d, J=8.1 Hz,2H), 6.87 (d, J=9.4 Hz, 2H), 6.71 (d, J=8.1 Hz, 2H), 6.00 (s, 2H), 5.85(s, 1H), 5.31 (s, 2H), 3.83 (s, 3H), 3.64 (s, 2H), 3.32 (s, 2H), 3.19(s, 2H), 2.39 (s, 3H), 1.50 (s, 2H), 1.44-1.37 (m, 2H), 0.87 (s, 6H). MS(ESI) m/z: 832.2 [M+H⁺].

Compound 310

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.2 Hz, 2H), 7.49 (d, J=8.3 Hz,2H), 7.41 (s, 1H), 6.91 (d, J=9.2 Hz, 2H), 6.72 (d, J=8.9 Hz, 1H), 6.20(s, 2H), 5.99 (s, 1H), 5.30 (s, 2H), 3.82 (s, 3H), 3.67 (s, 2H),3.48-3.33 (m, 8H), 2.38 (s, 3H), 1.66 (d, J=23.4 Hz, 4H). MS (ESI) m/z:834.2 [M+H⁺].

Compound 311

¹H NMR (400 MHz, CDCl3) δ 7.71 (d, J=8.5 Hz, 2H), 7.50 (d, J=8.5 Hz,2H), 6.93-6.89 (m, 2H), 6.72 (dd, J=9.0, 2.4 Hz, 1H), 5.84 (t, J=2.2 Hz,2H), 5.77 (s, 1H), 5.57 (s, 1H), 5.34 (t, J=2.2 Hz, 2H), 3.84 (s, 3H),3.66 (s, 2H), 3.23 (dd, J=12.8, 6.7 Hz, 2H), 2.41 (s, 3H), 1.46-1.24 (m,10H). MS (ESI) m/z: 818.1 [M+H⁺].

Compound 312

¹H NMR (400 MHz, CDCl3) δ 7.68 (d, J=8.5 Hz, 2H), 7.49 (d, J=8.5 Hz,2H), 6.88 (t, J=5.5 Hz, 2H), 6.70 (dd, J=9.0, 2.4 Hz, 1H), 6.10 (s, 2H),5.92 (s, 1H), 5.76 (s, 1H), 5.34 (d, J=1.9 Hz, 2H), 3.82 (s, 3H), 3.65(s, 2H), 3.24 (d, J=6.1 Hz, 2H), 2.39 (s, 3H), 1.71-1.63 (m, 2H), 1.58(s, 2H), 1.46-1.38 (m, 1H), 1.36-1.23 (m, 8H). MS (ESI) m/z: 832.1[M+H⁺].

Compound 313

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=7.6 Hz, 2H), 7.49 (d, J=7.6 Hz,2H), 6.87 (d, J=9.0 Hz, 2H), 6.70 (d, J=8.9 Hz, 1H), 5.87 (s, 2H), 5.71(s, 1H), 5.44 (s, 1H), 5.32 (s, 2H), 3.83 (s, 3H), 3.65 (s, 2H), 3.32(s, 2H), 2.40 (s, 3H), 1.40 (m, 14H). MS (ESI) m/z: 846.1 [M+H⁺].

Example 56 Synthesis of Compound 320

Intermediate 314

To a solution of propane-1,3-diamine (5.0 g, 67.5 mmol) in 100 mL ofMeOH: THF (2:1) was slowly added a solution of Boc₂O (7.4 g, 33.8 mmol)in THF (20 mL) at 0° C. The reaction mixture was allowed to stir at roomtemperature overnight. LCMS showed the reaction was completed, thesolvent was evaporated, water (100 mL) was added, and the mixture wasextracted with DCM (100 mL×3). The combined organic layers were washedwith brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated in vacuum to give tert-butyl (3-aminopropyl)carbamate as acolorless oil (1.8 g, yield: 32%). A mixture of tert-butyl(3-aminopropyl)carbamate (5.22 g, 30 mmol), 3-chloropropan-1-ol (11.28g, 120 mmol), K₂CO₃ (24.84 g, 180 mmol) and KI (996 mg, 6 mmol) in MeCN(300 mL) was stirred at 80° C. overnight. The solvent was evaporated andthe residue was washed with water (300 mL) and extracted with EtOAc (200mL×3). The combined organic layers were washed with brine, dried overNa₂SO₄, filtered and the filtrate was concentrated to give compound 314(6.5 g, 93%) as a yellow oil.

Intermediate 315

To a solution of compound 314 (6.5 g, 28 mmol) in dioxane (100 mL) wasadded NaOH (4 M, 21 mL) followed by CbzCl (4.76 g, 28 mmol) at 0° C. Theresulting mixture was stirred at rt overnight. The pH was adjusted topH=6 with HCl (1 N), and the mixture was extracted with EtOAc (100mL×3). The combined organic layers were washed with brine, dried overNa₂SO₄, filtered and the filtrate was concentrated. The residue waspurified by column chromatography on silica gel (eluent: PE/EtOAc: 8/1)to give tert-butyl (3-((3-hydroxypropyl)(2-oxo-2-phenyl-1λ²-ethyl)amino)propyl) carbamate as a colorless oil (7g, 74%). This was dissolved in a solution of HCl in Dioxane (4M, 5 ml)and stirred at rt for 2 h. The volatiles were removed to yield compound315 (2-((3-aminopropyl)(3-hydroxypropyl)amino)-1-phenyl-2λ²-ethan-1-one) which was used insubsequent reactions without further purification.

Intermediate 316

To a solution of indomethacin (366 mg, 0.93 mmol) in DMF (5 mL) wasadded compound 315 (300 mg, 1.12 mmol), HOBt (228 mg, 1.69 mmol), EDCI(323 mg, 1.69 mmol) and DIPEA (479 mg, 3.72 mmol) at 0° C. The resultingsolution was stirred at room temperature for 3.0 h. Then water was added(20 mL) and the mixture was extracted with EtOAc (30 mL×3). The combinedorganic layers were washed with brine, dried over Na₂SO₄, filtered andthe filtrate was concentrated to get the intermediate 316 (500 mg,yield: 89%) as a light yellow solid.

Intermediate 317

To a solution of 316 (500 mg, 0.83 mmol) in DCM (20 ml) at 0° C. wasadded Dess-Martin periodinane (385 mg, 0.91 mmol). The resultingsolution was stirred rt for 1.5 h, saturated Na₂S₂O₃ (10 mL) was addedand the mixture was stirred for 10 min, then washed with aqueous Na₂CO₃solution, extracted with DCM (30 mL×3). The combined organic layers werewashed with brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated to give compound 317 (490 mg, yield: 95%) as a yellowsolid.

Intermediate 318

To a solution of compound 317 (490 mg, 0.81 mmol) in MeOH (10 mL) wasadded 255 (352 mg, 0.81 mmol), and 0.5 mL of acetic acid. The resultingsolution was stirred at room temperature for 0.5 h. Then NaBH₃CN (256mg, 4.06 mmol) was added to the reaction mixture, which was stirred for2. h. Then water (20 mL) was added and the mixture was extracted withDCM (30 mL×3). The combined organic layers were washed with brine, driedover Na₂SO₄, filtered and the filtrate was concentrated to give thecrude product 318, which was purified by chromatography on silica gel(DCM/MeOH: 50/1) to give the product (360 mg, yield: 45%) as yellow oil.

Intermediate 319

318 (360 mg, 0.35 mmol) was dissolved in a solution of HBr in AcOH (40%,5 mL) and was stirred at room temperature for 2 h. The reaction wasconcentrated to give 319, which was used in the next step withoutpurification (290 mg, crude).

Compound 320

To a solution of 319 (290 mg, 0.32 mmol) in TFA (8 mL) was added Et₃SiH(68 mg, 0.59 mmol). The resulting mixture was stirred at 80° C. for 1.5h. The volatiles were removed and the residue was dissolved in NMP and(PPh₃)₂ReOCl₃ (Sigma-Aldrich, Order #370193, 1.0 eq) was added. Themixture was stirred at 80° C. overnight. The reaction was concentratedand the residue was purified by Prep-HPLC (Column: Acquity BEH C18,Waters Corp, A: water/B: MeCN) to give compound 320 as a light pinksolid (16 mg, yield: 6%)

¹H NMR (400 MHz, CDCl3) δ 8.29 (s, 1H), 7.67 (d, J=8.5 Hz, 2H), 7.49 (d,J=8.6 Hz, 2H), 6.97-6.82 (m, 2H), 6.74-6.57 (m, 2H), 4.76 (d, J=16.3 Hz,1H), 4.60-4.48 (m, 1H), 4.16-4.03 (m, 2H), 4.01-3.89 (m, 2H), 3.82 (s,3H), 3.75-3.61 (m, 3H), 3.56-3.43 (m, 1H), 3.38-3.05 (m, 5H), 2.98-2.65(m, 5H), 2.42-2.11 (m, 5H), 1.75-1.50 (m, 2H). MS (ESI) m/z: 848.1(M+H⁺).

Example 57 Synthesis of Compounds 328 and 329

Intermediate 322

To a solution of oxalyl chloride (3.1 g, 24.5 mmol) in dry DCM (200 mL)was added DMSO (2.9 mL, 40.8 mmol) at −60° C. The mixture was stirredfor 30 min. Then a solution of tert-butyl(R)-(1-hydroxypropan-2-yl)carbamate (3.5 g, 20 mmol) in DCM (20 mL) wasadded at −60° C. The resulting solution was stirred for another 30 min.Triethylamine (11.1 mL, 80 mmol) was added at −60° C., and the mixturewas stirred for 2 h at room temperature. Water (100 mL) was added andthe mixture was extracted with DCM (3×50 mL). The combined organiclayers were washed with brine, dried over Na₂SO₄, filtered and thefiltrate was concentrated to give the aldehyde 322 as a light yellow oil(3.2 g, 94%), which was used in the next step without furtherpurification.

Intermediate 323

To a solution of (4-methoxy-4-oxobutyl) triphenylphosphonium brominde(8.2 g, 18.5 mmol) in dry THF (150 mL) was added LiHMDS (1 M in THF, 20mL) dropwise. The resulting reaction was stirred at 0° C. for 30 min.Then the reaction was recooled to −60° C. A solution of compound 322(3.2 g, 18.5 mmol) in dry THF (50 mL) was added to the reaction. Thesolution was slowly warmed to RT and stirred overnight. The reaction wasquenched with NH₄Cl (aq.), extracted with EtOAc (100 mL×3). The combinedorganic layers were washed with brine, dried over Na₂SO₄, filtered andthe filtrate was concentrated in vacuo. The residue was purified bychromatography on silica gel (eluent: PE/EtOAc=10/1) to give thecompound 323 as a colorless oil (1.9 g, yield: 40%).

Intermediate 324

A mixture of compound 323 (1.9 g, 7.4 mmol) and Pd/C (200 mg) in MeOH(50 mL) was stirred under an atmosphere of H₂ overnight. The catalystwas filtered off, the filtrate was washed with MeOH (30 mL×3), andconcentrated to give 324 as a light yellow oil (1.9 g, yield: 99%) whichwas used without further purification.

Intermediate 325

To a solution of compound 324 (518 mg, 2 mmol) in dry THF (15 mL) wasadded LiAlH₄ (114 mg, 3 mmol) slowly at 0° C. The resulting reaction wasstirred at room temperature for another 2 h. The reaction was quenchedwith H₂O (15 ml), extracted with EtOAc (20 mL×3). The combined organiclayers were washed with brine, dried over Na₂SO₄, filtered and thefiltrate was concentrated. The residue was purified by chromatography onsilica gel (eluent: PE/EtOAc=4/1) to give the Boc-protected product as acolorless oil (270 mg, yield: 58%). This was dissolved in HCl (4M indioxane, 5 mL) and stirred at rt for 2 h, after which time LCMS showedthe reaction was completed. The reaction was concentrated to give 325,which was used without further purification (150 mg, yield: 99%).

Intermediate 326

326 was obtained by a sequence similar to the one described forintermediate 325, starting with (S)-(1-hydroxypropan-2-yl)carbamate.

Intermediate 327

To a solution of indomethacin (341 mg, 0.95 mmol) in DMF (5 mL) wasadded 325 (150 mg, 1.14 mmol), HOBt (230 mg, 1.71 mmol), EDCI (327 mg,1.71 mmol) and DIPEA (490 mg, 3.8 mmol) at 0° C. The resulting solutionwas stirred room temperature for 3.0 h. Then water was added (20 mL) andthe mixture was extracted with EtOAc (20 mL×3). The combined organiclayers were washed with brine, dried over Na₂SO₄, filtered and thefiltrate was concentrated to give 327 (300 g, yield: 67%) as a lightyellow solid.

Compound 328

Compound 328 was obtained from compound 327 by applying proceduressimilar to the ones described in Example 56 (compound 316 to compound320). The compound was purified by Prep-HPLC (Column: Acquity BEH C18,Waters Corp, A: water/B: MeCN) to give a light pink solid.

¹H NMR (400 MHz, CDCl₃) δ 7.68 (d, J=7.4 Hz, 2H), 7.50 (d, J=5.4 Hz,2H), 6.93-6.81 (m, 2H), 6.75-6.67 (m, 1H), 5.29 (d, J=8.4 Hz, 1H),4.69-4.52 (m, 2H), 4.14-3.95 (m, 3H), 3.94-3.85 (m, 1H), 3.83 (s, 3H),3.63 (s, 2H), 3.55-3.06 (m, 6H), 2.91-2.78 (m, 1H), 2.40 (d, J=1.8 Hz,3H), 1.60-1.52 (m, 1H), 1.38-1.15 (m, 7H), 1.05 (d, J=6.6 Hz, 3H). MS(ESI) m/z: 847.1 (M+H⁺).

Compound 329

Compound 329 was obtained by procedures similar to the ones describedfor compound 328, using intermediate 326.

¹H NMR (400 MHz, CDCl₃) δ 7.67 (d, J=8.5, 1.0 Hz, 2H), 7.50 (d, J=8.6Hz, 2H), 6.92-6.82 (m, 2H), 6.74-6.67 (m, 1H), 5.30 (d, J=8.5 Hz, 1H),4.67-4.52 (m, 2H), 4.14-3.95 (m, 3H), 3.95-3.75 (m, 4H), 3.63 (s, 2H),3.54-3.02 (m, 6H), 2.89-2.78 (m, 1H), 2.40 (d, J=1.8 Hz, 3H), 1.61-1.55(m, 1H), 1.48-1.13 (m, 7H), 1.05 (d, J=6.6 Hz, 3H). MS (ESI) m/z: 847.1(M+H⁺).

The following compounds 330-333 can also be prepared according to theprocedure given in Example 57 by replacing intermediates 325/326 withthe appropriate aminoalcohol.

Compound Y = 330

331

332

333

Example 58 Synthesis of Compound 341

Intermediate 335

To a solution of 334 (CAS No.: 25126-93-6, 3.45 g, 20.08 mmol) in THF(100 ml) was added lithium aluminum hydride (2.27 g, 60 mmol) at −78° C.After an hour the mixture was warmed to room temperature and stirredunder nitrogen for 4 h. The reaction was then quenched by the slowaddition of cold ethyl acetate. The white precipitate that formed wasthen filtered off and washed with cold ether to give the diol 335 3.2 g(100%) as a colorless oil.

Intermediate 336

To a solution of triphenylphosphine (2.0 g, 7.50 mmol) in THF (10 mL)was added N-chlorosuccinimide (1.0 g, 7.50 mmol). The reaction mixturewas allowed to stir at room temperature for 10 min, then 331 (400 mg,3.4 mmol) was added. The mixture was stirred overnight. Then water (20mL) was added. The mixture was extracted with EtOAc (30 mL×3), thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated to give the crude product,which was purified by chromatography on silica gel (EtOAc/hexanes) togive the dichloride 336 (260 mg yield: 50%) as a colorless oil.

Intermediate 337

To a solution of 336 (900 mg, 5.92 mmol) in DMF (15 mL) compound 255(850 mg, 1.97 mmol), K₂CO₃ (543 mg, 3.94 mmol) and KI (327 mg, 1.97mmol) were added. The mixture was heated to 80° C. overnight. Then water(50 mL) was added. The mixture was extracted with EtOAc (40 mL×3), thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated to give the crude productwhich was purified by chromatography on silica gel (DCM/MeOH=50/1) togive 337 (400 mg, yield: 37%) as yellow oil.

Intermediate 338

Compound 337 (400 mg, 0.72 mmol) was dissolved in a solution of NH₃ inMeOH (7M, 15 mL) and was heated to 110° C. in an autoclave overnight.The volatiles were removed and the crude product was purified bychromatography on silica gel (DCM/MeOH/H₂O=100:5:1) to give theintermediate 338 (240 mg, yield: 63%) as yellow oil.

Intermediate 339

To the solution of 338 (240 mg, 0.45 mmol) in DMF (10 mL) was addedindomethacin(2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3-yl)acetic acid,134 mg, 0.37 mmol), HOBt (92 mg, 0.68 mmol), EDCI (130 mg, 0.68 mmol)and DIPEA (190 mg, 1.48 mmol) at) 0° C.

The resulting solution was stirred at rt for 3 h. Water (20 mL) wasadded and the mixture was extracted with EtOAc (20 mL×4). The combinedorganic layers were washed with brine, dried over Na₂SO₄, filtered andthe filtrate was concentrated. Chromatography on silica gel(DCM:MeOH=50:1) afforded the product 339 (300 mg, yield: 93%) as a whitesolid.

Intermediate 340

To a solution of 339 (300 mg, 0.34 mmol) in TFA (2.0 mL) was addedEt₃SiH (75 mg, 0.68 mmol). The resulting mixture was stirred at 80° C.for 1.5 h. The reaction was concentrated to give 340 which was usedwithout further purification.

Compound 341

To a solution of 340 in NMP was added (PPh₃)₂ReOCl₃ (Sigma-Aldrich,Order #370193, 1.0eq) and the mixture was stirred at 80° C. overnight.The reaction was concentrated and the residue was purified by Prep-HPLC(Column: Acquity BEH C18, Waters Corp, A: water/B: MeCN) to givecompound 341 as a light pink solid (35 mg, yield: 10%).

¹H NMR (400 MHz, CDCl3) δ 7.67 (d, J=8.5 Hz, 2H), 7.50 (d, J=8.5 Hz,2H), 6.95-6.85 (m, 2H), 6.73 (dd, J=9.0, 2.5 Hz, 1H), 5.63 (t, J=5.5 Hz,1H), 5.49-5.36 (m, 1H), 5.18-5.08 (m, 1H), 4.70-4.53 (m, 2H), 4.13-4.03(m, 2H), 3.84 (s, 3H), 3.83-3.74 (m, 1H), 3.64 (s, 2H), 3.51-3.04 (m,8H), 2.86 (dd, J=13.5, 4.3 Hz, 1H), 2.36 (s, 3H), 2.34-2.21 (m, 2H),2.17-2.06 (m, 2H). MS (ESI) m/z: 831.1 (M+H⁺).

Example 59 Synthesis of Compound 346

Intermediate 342

To a solution of indomethacin(2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3-yl)acetic acid,3.57 g, 10 mmol) in dry DCM (50 mL) was added a solution of BBr₃ in DCM(1M, 13 mL) dropwise at −78° C. The resulting solution was stirred atroom temperature for another 1 h. LCMS showed the reaction wascompleted. Water (100 mL) was added to the reaction slowly, and themixture was extracted with DCM (70 mL×3). The combined organic layerswere washed with brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated to give the product (2.4 g, yield: 70%) as light yellowsolid.

Intermediate 343

To a solution of 342 (2.4 g, 4.86 mmol) and K₂CO₃ (520 mg, 4.86 mmol) inDMF (30 mL) was added BnBr (0.3 mL) dropwise. The mixture was stirred atroom temperature for 1.0 h, then water (50 mL) was added and the mixturewas extracted with EtOAc (50 mL×3). The combined organic layers werewashed with brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated. The residue was purified by chromatography on silica gel(PE/EA=2/1) to give the benzyl ester 343 (2.5 g yield: 83%) as a whitesolid.

Intermediate 344

To a solution of 343 (800 mg, 2.54 mmol) and PPh₃ (1.45 g, 7.62 mmol) inTHF (30 ml) was added CD₃OD (135 mg, 5.08 mmol) under N₂ at 0° C. Themixture was stirred for 20 min, and then DEAD (964 mg, 7.62 mmol) wasadded dropwise at 0° C. The mixture was stirred at room temperatureovernight. Then water (30 mL) was added and the mixture was extractedwith EtOAc (40 mL×3). The combined organic layers were washed withbrine, dried over Na₂SO₄, filtered and the filtrate was concentrated.The residue was purified by chromatography on silica gel (PE/EA=10/1) toget the product 344 (450 mg, yield: 40%) as a white solid.

Intermediate 345

A mixture of compound 344 (300 mg, 0.66 mmol) and Pd/C (60 mg) in THF(20 mL) was stirred under H₂ for 3 h. The catalyst was filtered off, thefiltrate was washed with THF (10 mL×3), and concentrated to give2-(1-(4-chlorobenzoyl)-5-(methoxy-d₃)-2-methyl-1H-indol-3-yl)acetic acid345 (240 mg, 98%) which was used without further purification.

Compound 346 was synthesized from2-(1-(4-chlorobenzoyl)-5-(methoxy-d₃)-2-methyl-1H-indol-3-yl)acetic acid345 by a procedure similar to the one described in Example 57, using6-aminohexan-1-ol instead of intermediate 315. ¹H NMR (400 MHz, CDCl3) δ7.68 (d, J=8.5 Hz, 2H), 7.50 (d, J=8.5 Hz, 2H), 6.93-6.79 (m, 2H), 6.71(dd, J=9.0, 2.5 Hz, 1H), 5.60 (t, J=5.7 Hz, 1H), 4.67-4.51 (m, 2H),4.17-4.01 (m, 2H), 3.97-3.81 (m, 1H), 3.65 (s, 2H), 3.53-3.41 (m, 1H),3.39-3.07 (m, 6H), 2.85 (dd, J=13.4, 4.2 Hz, 1H), 2.40 (s, 3H),1.80-1.63 (m, 2H), 1.50-1.38 (m, 2H), 1.36-1.20 (m, 4H). MS (ESI) m/z:836.0 (M+H⁺).

Example 60 Synthesis of Compound 351

2-(1-(4-Chlorobenzoyl)-5-methoxy-2-(trifluoromethyl)-1H-indol-3-yl)aceticacid, 350, can be synthesized as described by Anna L. Blobaum, Md.Jashim Uddin, Andrew S. Felts, Brenda C. Crews, Carol A. Rouzer, andLawrence J. Marnett, ACS Med. Chem. Lett., 4, 486-490, (2013).Alternatively, 350 can be synthesized as outlined below.

Intermediate 347

2-(5-methoxy-1H-indol-3-yl)acetic acid (5 g, 24.4 mmol) and BOP-Cl (7.4g, 29 mmol) were suspended in DCM (100 mL), and Et₃N (5.4 g, 53.7 mmol)was slowly added. The resulting solution was stirred for 30 min. Benzylalcohol (2.9 g, 26.8 mmol) was added and the mixture was stirred for 12h. DCM (100 mL) was added, the layers were separated and the organiclayer was washed with water (200 mL) and brine. The organic layer wasconcentrated and purified by chromatography on silica gel (eluent:PE/EtOAc=2/1) to give 347 as a white solid (6.1 g, 85%).

Intermediate 348

A mixture of compound 347 (4.7 g, 15.82 mmol) and Togni's reagent(1-Trifluoromethyl-1,2-benziodoxol-3-(1H)-one, 6.5 g, 20.56 mmol) inMeOH (100 mL) was degassed for 15 min, then CuOAc (1.4 g, 11 mmol) wasadded. The resulting mixture was stirred at rt overnight. The solventwas removed; water (100 mL) was added, extracted with EtOAc (100 mL×2).The combined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated. The residue was purified bychromatography on silica gel (eluent: PE/EtOAc=5/1) to give the productas a white solid (4 g, 70%).

Intermediate 349

To a solution of compound 348 (4 g, 11 mmol) in dry DMF (50 mL) wasadded NaH (750 mg, 18.7 mmol). The reaction was stirred for another 30min, and then 4-chlorobenzoyl chloride (2.9 g, 16.5 mmol) was added. Theresulting mixture was stirred at room temperature overnight. Thereaction was quenched by addition of sat′d aq, NH₄Cl solution (100 mL),extracted with EtOAc (100 mL×3). The combined organic layers were washedbrine, dried over Na₂SO₄, filtered and the filtrate was concentrated.The residue was purified by chromatography on silica gel (eluent:PE/EtOAc=2/1) to give the product as a yellow solid (3.3 g, 50%).

Intermediate 350

A mixture of compound 349 (3.3 g, 6.5 mmol) and Pd/C (1 g) in THF (30mL) was stirred at 40° C. for 1 h under H₂ atmosphere. The catalyst wasfiltered off and washed with MeOH (30 mL×3). The filtrate wasconcentrated in vacuo to give the crude product which was purified bychromatography on silica gel (eluent: DCM/MeOH=40/1) to give2-(1-(4-chlorobenzoyl)-5-methoxy-2-(trifluoromethyl)-1H-indol-3-yl)acetic acid, 350, as a solid (1.6 g, yield: 60%).

Compound 351 was synthesized from2-(1-(4-chlorobenzoyl)-5-methoxy-2-(trifluoromethyl)-1H-indol-3-yl)acetic acid, 350, by a procedure similar to the one described in Example42.

¹H NMR (400 MHz, CDCl3) δ 7.83 (d, J=8.7 Hz, 2H), 7.56 (d, J=7.7 Hz,2H), 7.15 (s, 1H), 6.91 (d, J=9.0 Hz, 1H), 6.73 (d, J=9.2 Hz, 1H), 5.60(s, 1H), 4.74-4.54 (m, 2H), 4.23-4.04 (m, 2H), 4.00-3.74 (m, 6H), 3.49(s, 1H), 3.42-3.06 (m, 6H), 2.89 (s, 1H), 1.69-1.44 (m, 4H), 1.42-1.25(m, 4H). MS (ESI) m/z: 887.1 (M+H⁺).

The following compound 352 can also be prepared according to theprocedure given in Example 60.

¹H NMR (400 MHz, CDCl3) δ 7.80 (d, J=8.5 Hz, 2H), 7.53 (d, J=8.4 Hz,2H), 7.11 (d, J=2.2 Hz, 1H), 6.88 (dd, J=9.2, 2.3 Hz, 1H), 6.71 (d,J=9.2 Hz, 1H), 5.62 (s, 1H), 4.76-4.51 (m, 1H), 4.20-4.02 (m, 2H),4.02-3.79 (m, 6H), 3.60-3.42 (m, 1H), 3.41-3.09 (m, 7H), 2.86 (dd,J=13.4, 4.2 Hz, 1H), 1.60-1.38 (m, 2H), 1.41-1.09 (m, 8H). MS (ESI) m/z:901.1 (M+H⁺).

Example 61 Synthesis of Compound 356

Synthesis of intermediate 353

A mixture of compound 255 (1.3 g, 3 mmol), 3-chloropropanoic acid (1.3g, 12 mmol) and TEA (1.52 g, 15 mmol) in MeCN (30 mL) was stirred at 80°C. overnight. LCMS showed the reaction was completed. The reaction wasconcentrated in vacuo, washed with water (50 mL) and extracted withEtOAc (50 mL×3). The combined organic layers were washed brine, driedover Na₂SO₄, filtered and the filtrate was concentrated to give 353 as ayellow oil (1.4 g, 92%).

Synthesis of intermediate 354

To a solution of 353 (1.4 g, 2.77 mmol) in DMF (20 mL) were addedcompound 296 (Example 52, 950 mg, 2.77 mmol), HOBt (560 mg, 4.15 mmol),EDCI (792 mg, 4.15 mmol) and DIPEA (1.8 g, 13.8 mmol) at 0° C. Theresulting solution was stirred rt overnight. Then water (40 mL) wasadded and the mixture was extracted with EtOAc (50 mL×4). The combinedorganic layers were washed with brine, dried over Na₂SO₄, filtered andthe filtrate was concentrated. The residue was purified bychromatography on silica gel (eluent: DCM/MeOH=40/1) to give the product(1 g, 43%) as light yellow solid.

Synthesis of intermediate 355

At 0° C., to a solution of compound 3 (300 mg, 0.36 mmol) in TFA (2 mL)was added anisole (0.1 mL). MeSO₃H (1 mL) was added dropwise. Theresulting reaction was stirred at room temperature for another 1 h. WhenLCMS showed completion of the reaction, the mixture was concentrated togive the crude product 355 (310 mg) which was used without furtherpurification.

A solution of 355 (310 mg) and (PPh₃)₂ReOCl₃ (Sigma-Aldrich, Order#370193, 300 mg, 0.36 mmol) was stirred at 80° C. for 16 h. The reactionwas concentrated and purified by Prep-HPLC (Column: Acquity BEH C18,Waters Corp, A: water/B: MeCN) to give 356 as a light pink solid (30 mg,yield: 10%).

¹H NMR (400 MHz, CDCl3) δ 7.71-7.59 (m, 2H), 7.55-7.43 (m, 2H), 6.94 (s,1H), 6.86 (d, J=9.0 Hz, 1H), 6.67 (d, J=9.0 Hz, 1H), 5.90 (s, 1H), 4.61(d, J=16.5 Hz, 1H), 4.51 (dd, J=11.6, 5.1 Hz, 1H), 4.07 (dd, J=10.5, 3.9Hz, 1H), 4.03-3.74 (m, 6H), 3.63-3.36 (m, 3H), 3.29-3.04 (m, 3H),2.98-2.78 (m, 3H), 2.65 (t, J=6.6 Hz, 2H), 2.36 (s, 3H). MS (ESI) m/z:790.8 (M+H+)

Example 62 Synthesis of Compound 359

Synthesis of intermediate 357

A mixture of 255 (868 mg, 2 mmol), methyl 6-bromohexanoate (1.3 g, 6mmol), K₂CO₃ (1.3 g, 10 mmol) and KI (500 mg, 3 mmol) in MeCN (50 mL)was stirred at 80° C. overnight. Then water (40 mL) was added and themixture was extracted with EtOAc (50 mL×4). The combined organic layerswere washed brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated to give 357 as a white solid (1.1 g, yield: 92%).

Synthesis of intermediate 358

A mixture of 357 (1.1 g, 1.95 mmol) and NaOH (156 mg, 3.9 mmol) inTHF/MeOH/H₂O (10 mL/10 mL/4 mL) was stirred at rt. Once LCMS showed thereaction to be completed, the pH was adjusted to 4 and the mixture wasextracted with EtOAc (30 mL×3). The combined organic layers were washedwith brine, dried over Na₂SO₄, filtered and the filtrate wasconcentrated to give 358 as light yellow solid (850 mg, 79%).

Compound 359 was synthesized from intermediate 358 by a proceduresimilar to the one described in Example 61.

¹H NMR (400 MHz, CDCl3) δ 7.66 (d, J=8.4 Hz, 2H), 7.48 (d, J=8.3 Hz,2H), 6.98 (d, J=16.3 Hz, 1H), 6.86 (d, J=9.0 Hz, 1H), 6.68 (d, J=9.0 Hz,1H), 5.53 (s, 1H), 4.82-4.33 (m, 2H), 4.07 (dd, J=21.9, 13.1 Hz, 2H),3.98-3.72 (m, 4H), 3.58-3.05 (m, 7H), 2.88 (dt, J=13.7, 5.4 Hz, 3H),2.45 (d, J=63.8 Hz, 3H), 2.16 (dd, J=17.8, 10.6 Hz, 2H), 1.95-1.56 (m,4H), 1.38 (dd, J=15.1, 7.5 Hz, 2H). MS (ESI) m/z: 833.1 (M+H⁺).

Example 63

The ^(99m)Tc complex 360 is prepared as described in Example 62 using asthe last step a ligand exchange reaction employing^(99m)Tc-glucoheptonate as described in e.g.: Ono, M., et al., ACS Chem.Neurosci. 1, 598, (2010) and Ono, M., et al., Bioorg. Med. Chem. Lett.,20, 5743-5748, (2010).

Example 64 Synthesis of Compound 365

Intermediate 361

To a solution of 4-fluorophenyl methyl ketone (8.28 g, 60 mmol) in dryTHF (200 mL) was added NaH (3.12 g, 78 mmol), followed by dimethyloxalate (7.8 g, 66 mmol). The resulting solution was stirred at 70° C.overnight. LCMS showed the reaction was completed. The reaction wasquenched by addition of sat′d aq. NH₄Cl solution and extracted withEtOAc (200 mL×3). The combined organic layers washed with brine, driedover Na₂SO₄, filtered and the filtrate was concentrated to give thecrude product which was purified by chromatography on silica gel(eluent: PE/EtOAc=8/1) to give 361 (10 g, yield: 75%) as a colorlessoil.

Intermediate 362

A solution of 361 (10 g, 44.6 mmol) and 4-hydrazinylbenzenesulfonamide(8.34 g, 44.6 mmol) in MeOH (150 mL) was stirred at 70° C. overnight.LCMS showed the reaction was completed. The reaction was concentrated,washed with water (150 mL) and extracted with EtOAc (150 mL×3). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated to give 362 (15 g, 90%) as awhite solid.

Intermediate 363

To a solution of 362 (15 g, 40 mmol) in dry THF (200 mL) at 0° C. wasslowly added LiAlH₄ (1.82 g, 48 mmol). The resulting reaction wasstirred at room temperature for 1 h. LCMS showed the reaction wascompleted. The reaction was quenched with 2 mL of water at 0° C., then150 mL water was added thereto. The mixture was extracted with EtOAc(150 mL×3) and the combined organic layers were washed with brine, driedover Na₂SO₄, filtered and the filtrate was concentrated to give thecrude product, which was purified by chromatography on silica gel(eluent: DCM/MeOH=40/1) to give 363 (9.5 g, yield: 68%).

Intermediate 364

To a solution of compound 4 (500 mg, 1.44 mmol) in dry THF (20 mL) at 0°C. was added MsCl (248 mg, 2.16 mmol), followed by TEA (363 mg, 3.6mmol). The resulting mixture was stirred at room temperature for another1 h. The reaction was judged complete by TLC and H₂O (20 mL) was added.The mixture was extracted with EtOAc (30 mL×3). The combined organiclayers were washed with brine, dried over Na₂SO₄, filtered and thefiltrate was concentrated to give a colorless oil. This was dissolved ina solution of NH₃ in MeOH (7 M, 5 mL) and stirred at room temperaturefor 1 h. LCMS showed the reaction was completed. The reaction wasconcentrated in vacuo to give 364 (450 mg, yield: 93%).

To a solution of 364 (450 mg, 1.3 mmol) in DMF (10 mL) acid 241 (580 mg,1.5 mmol), HOBt (350 mg, 2.6 mmol), EDCI (497 mg, 2.6 mmol) and DIPEA(671 mg, 5.2 mmol) were added at 0° C. The resulting solution wasstirred room temperature overnight. The reaction was quenched withsaturated NaHCO₃ solution (30 ml) and extracted with ethyl acetate (2×50ml). The combined organic extracts were washed with brine (30 ml), driedover Na₂SO₄, concentrated, and purified by prep-HPLC (Column: AcquityBEH C18, Waters Corp, solvent A: water/solvent B: MeCN) to give thecompound 365 (90 mg, yield: 10%).

¹H NMR (400 MHz, CDCl3) δ 7.65 (d, J=8.0 Hz, 2H), 7.21-6.89 (m, 7H),6.55 (s, 1H), 6.07 (d, J=39.8 Hz, 4H), 5.36 (s, 2H), 4.62 (d, J=4.9 Hz,2H). MS (ESI) m/z: 709.0 (M+H⁺)

Example 65 Synthesis of Compound 366

Compound 366 is prepared from compound 364 (Example 64) by a proceduresimilar to the one described in Example 41.

Example 66 Synthesis of Compound 367

Compound 367 is prepared from compound 364 (Example 64) and compound 358(Example 62) using a procedure similar to the one described in Example61.

Example 67 Synthesis of Compound 368

The ^(99m)Tc complex 368 is prepared as described in Example 66 using asthe last step a ligand exchange reaction employing^(99m)Tc-glucoheptonate as described in e.g.: Ono, M., et al., ACS Chem.Neurosci. 1, 598, (2010) and Ono, M., et al., Bioorg. Med. Chem. Lett.,20, 5743-5748,

Example 68 Synthesis of Compound 371

Intermediate 369

To a solution of 296 (170 mg, 0.5 mmol) and picolinaldehyde (54 mg, 0.5mmol) in DCM (15 ml) was added AcOH (0.2 ml). The mixture was stirred atroom temperature for 30 min, NaBH(OAc)₃ (422 mg, 2.0 mmol) was added,the mixture was stirred at room temperature overnight, then water (20mL) was added and the mixture was extracted with EtOAc (20 mL×3). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and the filtrate was concentrated. The residue was purified bychromatography on silica gel (DCM/MeOH=10/1) to give 369 (60 mg yield:28%).

Intermediate 370

To a solution of 369 (120 mg, 0.3 mmol) and 2-oxoacetic acid (50% inwater, 133 mg. 0.9 mmol) in MeOH (10 mL) was added AcOH (0.1 mL). Themixture was stirred at room temperature for 30 min, then NaCNBH₃ (93 mg,1.5 mmol) was added and the mixture was stirred at room temperatureovernight. Water (20 mL) was added and the mixture was extracted withEtOAc (20 mL×3). The combined organic layers were washed with brine,dried over Na₂SO₄, filtered and the filtrate was concentrated. Theresidue was purified by chromatography on silica gel (DCM/MeOH=3/1) togive 370 (30 mg yield: 20%).

To a solution of compound 370 (30 mg, 0.065 mmol) in MeOH (3 mL) wasadded Re(CO)₅Cl (36 mg, 0.1 mmol). The resulting mixture was refluxedovernight. LCMS showed the reaction to be complete. The reaction mixturewas concentrated in vacuo to give the crude product, which was purifiedby Prep-HPLC (Column: Acquity BEH C18, Waters Corp.) to give 371 (10 mgyield: 20%) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.80 (d, J=5.3 Hz, 1H), 8.20 (t, J=7.6 Hz,1H), 7.79 (d, J=7.9 Hz, 1H), 7.72 (d, J=8.4 Hz, 2H), 7.63 (dd, J=18.1,7.5 Hz, 3H), 7.25 (d, J=2.1 Hz, 1H), 6.91 (d, J=9.0 Hz, 1H), 6.74 (dd,J=9.0, 2.1 Hz, 1H), 4.94 (dd, J=35.8, 15.8 Hz, 2H), 4.14 (d, J=16.7 Hz,1H), 3.79 (s, 3H), 3.72-3.48 (m, 3H), 3.25-3.09 (m, 2H), 2.33 (s, 3H).MS (ESI) m/z: 762.0 [M+H⁺].

Clinical and Screening Examples

Example A “Pain Scans” to Localize Site(s) of Inflammation

A patient with an undiagnosed cause of pain, or a cause of pain whichcannot be localized to a site of pathology, is scheduled for a “painscan.” The patient refrains from drinking or eating for at least eighthours prior to the pain scan. A compound of the invention isadministered to the patient either orally or parenterally. After anappropriate period of time determined by the pharmacokinetics of thecompound of the invention, during which the compound of the inventionbinds to cyclooxygenase, the patient is scanned with the appropriatemodality to determine the locus or loci of the greatest concentration ofthe compound. The loci are imaged and viewed or photographed asappropriate. Scans of the patient can be repeated at various intervalsafter ingestion or injection of the compound of the invention, forexample, at two hours, three hours, and four hours after ingestion orinjection. The scan findings are correlated with the patient's medicalhistory, physical examination and other information to assist indiagnosis of the etiology of the pain and determine appropriatetreatment.

Example B “Tumor Scans” to Localize Site(s) of Tumor(s)

A patient to be screened for presence of a tumor is scheduled for a “COXscan.” The patient refrains from drinking or eating for at least eighthours prior to the COX scan. A compound of the invention is administeredto the patient either orally or parenterally. After an appropriateperiod of time determined by the pharmacokinetics of the compound of theinvention, during which the compound of the invention binds tocyclooxygenase, the patient is scanned with the appropriate modality todetermine the locus or loci of the greatest concentration of thecompound. The loci are imaged and viewed or photographed as appropriate.Scans of the patient can be repeated at various intervals afteringestion or injection of the compound of the invention, for example, attwo hours, three hours, and four hours after ingestion or injection. Thescan findings are correlated with the patient's medical history,physical examination and other information to assist in diagnosis of thepresence and/or location of the tumor and determine appropriatetreatment.

Example C Scans to Screen for Asymptomatic Infections or LocalizedInfections

A patient to be screened for an asymptomatic infection, or to have thesite of a localized infection identified, is scheduled for a “COX scan.”The patient refrains from drinking or eating for at least eight hoursprior to the COX scan. A compound of the invention is administered tothe patient either orally or parenterally. After an appropriate periodof time determined by the pharmacokinetics of the compound of theinvention, during which the compound of the invention binds tocyclooxygenase, the patient is scanned with the appropriate modality todetermine the locus or loci of the greatest concentration of thecompound. The loci are imaged and viewed or photographed as appropriate.Scans of the patient can be repeated at various intervals afteringestion or injection of the compound of the invention, for example, attwo hours, three hours, and four hours after ingestion or injection. Thescan findings are correlated with the patient's medical history,physical examination and other information to assist in diagnosis of thepresence and/or location of an infection, and to determine appropriatetreatment.

Example D Scans to Screen Candidate Compounds for Imaging

Animal models can be used to test the conjugates and compounds of theinvention for their suitability for clinical use. Animal models of pain(and inflammation related to pain), of infection, and of cancer are wellknown. See, for example, Handbook of Laboratory Animal Sciense, SecondEdition: Animal Models, Volume 2 (Jann Hau, Gerald L. Van Hoosier Jr.,editors), Boca Raton: CRC Press, 2003; Animal Models for the Study ofHuman Disease (P. Michael Conn, editor), San Diego: Academic Press,2013.

A suitable animal model (for pain, for cancer, or for infection) isselected and the appropriate pathology is induced. The site of theinduced pain, inflammation, infection, or tumor is recorded by theinvestigator. One or more candidate conjugates or compounds of theinvention is administered to the animal, either by oral gavage orparenterally. After an appropriate period of time determined by thepharmacokinetics of the compound of the invention, during which thecompound of the invention binds to cyclooxygenase, the animal is scannedwith the appropriate modality to determine the locus or loci of thegreatest concentration of the compound. The location(s) indicated by thescan are compared with the known site or sites at which the pathologywas induced, for evaluation of the effectiveness of the conjugate foraccumulating at the site of pathology.

The carrageenan induced rat paw edema assay can be used as an exemplarymodel for inflammation; see Shalini, V. et al., Molecular Immunology66:229-239 (2015); see also Winter, C. et al., Proc. Soc. Exp. Biol.Med. 111:544-547 (1962). Briefly, acute inflammation is induced byaponeurosis injection of 0.1 ml of 1% carrageenan in 0.9% saline.Additional information regarding model assays is described in Guay etal., J. Biol. Chem. 279:24866-24872 (2004); Nantel et al., BritishJournal of Pharmacology 128:853-859 (1999); Siebert et al., Proc. Natl.Acad. Sci. USA 91:12013-12017 (1994); de Vries et al., J Nucl. Med.44:1700-1706 (2003); and Uddin et al., Cancer Prev. Res. 4:1536-1545(2011).

The animal is then imaged using the appropriate modality, for example,scintigraphic imaging or SPECT imaging. Exemplary imaging methods thatcan be used are described in Pacelli et al., J. Label. Compd.Radiopharm. 57:317-322 (2014); de Vries et al., J Nucl. Med.44:1700-1706 (2003); and Tietz et al., Current Medicinal Chemistry, 20,4350-4369 (2013).

Example E COX Inhibition Assay

A variety of assays can be used to evaluate inhibition of compounds tocyclooxygenase (COX). Compounds in the present invention displayinhibition of cyclooxygenase in the following assays.

Cell Culture:

RAW264.7 murine macrophages were obtained from the Cell Bank of ShanghaiInstitute of Biochemistry and Cell Biology, Chinese Academy of Sciences(Shanghai, China) and cultured in Dulbecco's modified Eagle's mediumcontaining 100 U/ml penicillin and 100 μm/ml streptomycin at 37° C. in5% CO₂.

Cell-Based COX-2 Assay:

RAW264.7 cells were plated at a density of 2.5×105/m1 cells in a 96-wellplate with 0. 1 ml of culture medium per well and cultured overnight.The cells were pre-incubated for 30 min with various doses of compoundsand stimulated for 7 h with 1m/m1 LPS and 10 U/ml IFN-g. The cellculture supernatants were collected immediately following treatment andcentrifuged at 1,000 rpm for 5 min to remove the particulate matter.PGE2 was determined using a Prostaglandin E2 assay kit (catalog no.62P2APEB; Cisbio Co.,). 10 μL of cell supernatant was transferred to a384-well low volume plate (e.g. Corning® 3544), 5 μL of PGE2-d2 wasadded, followed by 5 μL anti-PGE2 Cryptate as a negative control.Replace the standard by 10 μL of diluent and PGE2-d2 by 5 μL ofreconstitution buffer, Cal0 (for positive control), replace the standardby 10 μL of diluent. Incubate at 4° C. overnight. After centrifuging at1,000 rpm for 1 min, the dual fluorescence emissions of 615 and 665 nmwith a 320 nm excitation were measured using an Envision plate reader(Perkin Elmer, Shelton, Conn.). The results are expressed as the ratioof 665 nm/615 nm emissions.

COX-1/-2 Enzyme Assay:

The ability of compounds to inhibit ovine COX-1 and human COX-2 wasdetermined using a commercially available enzyme immunoassay (EIA) kit(catalog no. 701090 (COX-1); 701080 (COX-2) Cayman Chemical Co., AnnArbor, Mich., USA) according to the manufacturer's protocol. COXcatalyzes the first step in the biosynthesis of AA to PGH2. PGF2α,produced from PGH2 by reduction with stannous chloride, was measured byEIA (ACE™ competitive EIA, Cayman Chemical, Ann Arbor, Mich., USA).Briefly, to a series of supplied reaction buffer solutions [960 μl 0.1 MTris-HCl (pH 8.0) containing 5 mM EDTA and 2 mM phenol] with eitherCOX-1 or COX-2 (10 μl) enzyme in the presence of heme (10 μl), 10 μl ofvarious concentrations of test drug solutions were added. Thesesolutions were incubated for 15 min at 37° C. and subsequently 10 μl AAsolution (100 μM) was added. The COX reaction was stopped by theaddition of 30 μl stannus chloride after 2 min, mixed immediately,supernatants were 2000 fold diluted. The produced PGF2α was measured byEIA. This assay is based on the competition between PGs and aPG-acetylcholinesterase conjugate (PG tracer) for a limited amount of PGantiserum. The amount of PG tracer that is able to bind to the PGantiserum is inversely proportional to the concentration of PGs in thewells since the concentration of the PG tracer is held at a constantwhile the concentration of PGs varies. The specific antiserum-PG complexbound to a mouse anti-rabbit IgG that had been previously attached tothe well. The plate was washed to remove any unbound reagents and 200 μlEllman's reagent (5,5′-dithiobis-(2-nitrobenzoic acid), which containsthe substrate to acetylcholine esterase, was added to the well. Theproduct of this enzymatic reaction generates a distinct yellow colorthat absorbs at 406 nm. The intensity of this color, determined byspectrophotometry, is proportional to the amount of PG tracer bound tothe well, which is inversely proportional to the amount of PGs presentin the well during the incubation. Percent inhibition was calculated bythe comparison of the compounds treated to the various controlincubations.

Dose-response curves were generated using XLFit (IDBS, Surrey, UK) orPrism (GraphPad Software, La Jolla, Calif., US) to calculate IC₅₀ valuesfor each compound tested.

Representative results for COX-2 inhibition are provided in Table 3below. IC₅₀ values are given in micromolar units.

TABLE 3 COX-2 IC₅₀ (μM) Compound enzyme cell 23 12.0 — 27 0.6 12.7 351.6 — 39 1.1 — 178 0.2 1.8 181 1.0 1.5 182 0.8 0.6 183 0.4 0.7 185 1.20.5 204 0.2 0.9 207 0.6 0.8 208 0.3 0.6 209 2.4 0.7 210 0.3 0.6 211 2.21.6 219 4.0 2.1 234 1.7 3.7 237 4.5 >1 242 0.3 0.4 243 0.5 0.7 244 >301.5 245 >30 0.3 246 0.1 0.2 247 0.2 2.6 248 0.2 >30 249 1.1 4.4 250 0.20.3 251 0.3 >30 260 0.3 0.2 261 0.8 1.8 262 0.9 1.7 263 0.3 0.3 264 0.30.1 265 2.1 0.1 266 0.6 0.2 267 >30 0.3 268 >30 >10 269 0.5 0.9 275 0.70.2 297 0.5 0.8 301 >30 >1 303 >30 0.9 304 >30 0.9 305 0.3 0.1 306 0.70.2 307 >30 0.5 308 0.9 0.2 309 0.5 0.3 310 >30 0.2 311 — 0.4 312 0.10.4 313 — 0.2 320 — 5.0 328 0.6 0.1 329 0.6 0.1 341 0.8 0.3 346 0.3 0.2351 >30 0.4 352 >30 0.3 356 — 1.6 359 0.6 0.2 365 7.9 1.1 367 4.5 >10371 0.1 2.1

Representative results for COX-1 inhibition are provided in Table 4below. IC₅₀ values are given in micromolar units.

TABLE 4 COX-1 IC₅₀ (μM) Compound enzyme 242 >30 246 >30 260 >30 264 >30275 >30 305 >30 306 >30 275 >30 369 >30

Example F: Pharmacokinetic Data

In vitro data of metabolic stability and protein binding as well as invivo pharmacokinetic data for the conjugates of the invention can begenerated using the techniques disclosed in Silber, B. M. et al., Pharm.Res. 30(4):932-950 (2013), which is hereby incorporated by reference inits entirety. Various biological, pharmacokinetic and other propertiesof the conjugates, including hepatic microsomal stability, determinationof metabolites, binding to proteins such as plasma protein binding, andin vivo studies, including single-dose and multi-dose pharmacokineticstudies, are determined using the protocols described in thatpublication and the publications cited therein.

Exemplary Embodiments

The invention is further described by the following exemplaryembodiments. The features of each of the embodiments are combinable withany of the other embodiments where appropriate and practical.

Embodiment 1

A conjugate comprising: a non-steroidal anti-inflammatory drug (NSAID)or a residue of a NSAID bonded or complexed to an imaging moiety whichis a radioactive agent, wherein the radioactive agent is selected fromthe group consisting of a gamma-ray emitter and an X-ray emitter; or apharmaceutically acceptable salt thereof.

Embodiment 2

The conjugate of embodiment 1, wherein said radioactive agent is^(99m)Tc.

Embodiment 3

The conjugate of embodiment 1, wherein the NSAID or the residue of aNSAID and imaging moiety are bonded or complexed via a linker selectedfrom the group consisting of an optionally substituted C₁-C₃₀hydrocarbylene group and an optionally substituted C₃-C₃₀heterohydrocarbylene group.

Embodiment 4

The conjugate of any of embodiments 1-3, wherein the NSAID or theresidue of a NSAID is selected from the group consisting ofacetylsalicylic acid, diflunisal, salsalate, choline magnesiumtrisalicylate, ibuprofen, dexibuprofen, naproxen, fenoprofen,ketoprofen, dexketoprofen, flurbiprofen, oxaprozin, loxoprofen,indomethacin, tolmetin, sulindac, etodolac, ketorolac, diclofenac,aceclofenac, nabumetone, piroxicam, meloxicam, tenoxicam, droxicam,lornoxicam, isoxicam, mefenamic acid, meclofenamic acid, flufenamicacid, tolfenamic acid, celecoxib, rofecoxib, valdecoxib, parecoxib,lumiracoxib, etoricoxib, firocoxib, nimesulide, and licofelone, and aresidue of any of the foregoing compounds.

Embodiment 5

The conjugate of any of embodiments 1-3, wherein the optionallysubstituted C₃-C₃₀ heterohydrocarbylene group is

where R¹ and R² are independently selected from hydrogen, C₁-C₄ alkyl,C₁-C₄ alkyl substituted with hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆cycloalkyl; or R¹ and R² together with the carbon to which they areattached form a C₃-C₈ cycloalkyl ring; and n is an integer selected from0 to 4, inclusive.

Embodiment 6

The conjugate of embodiment 5, wherein R¹ and R² are methyl.

Embodiment 7

The conjugate of embodiment 5, wherein the linking group is

Embodiment 8

The conjugate of any of embodiments 1-3, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID is selected fromthe group consisting of naproxen, ketorolac, a residue of naproxen, anda residue of ketorolac.

Embodiment 9

The conjugate of embodiment 1, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID bonded orcomplexed to an imaging moiety which is a radioactive agent is:

wherein Tc is ^(99m)Tc; or a pharmaceutically acceptable salt thereof.

Embodiment 10

The conjugate of embodiment 1, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID bonded orcomplexed to an imaging moiety which is a radioactive agent is:

wherein Tc is ^(99m)Tc; or a pharmaceutically acceptable salt thereof.

Embodiment 11

A pharmaceutical composition comprising the conjugate of any ofembodiments 1-3, 9, or 10, and a pharmaceutically acceptable excipient.

Embodiment 12

A method of imaging a site of pathology in a subject, comprising: a)administering a conjugate comprising a non-steroidal anti-inflammatorydrug (NSAID) or a residue of a NSAID bonded or complexed to an imagingmoiety which is a radioactive agent, wherein the radioactive agent isselected from the group consisting of a gamma-ray emitter and an X-rayemitter; or a pharmaceutically acceptable salt of said conjugate; or apharmaceutical composition comprising said conjugate or apharmaceutically acceptable salt of said conjugate and apharmaceutically acceptable excipient; and b) generating an image of thesubject or an image of a portion of the subject.

Embodiment 13

The method of embodiment 12, wherein said radioactive agent is ^(99m)Tc.

Embodiment 14

The method of embodiment 12, wherein the NSAID or the residue of a NSAIDand imaging moiety are bonded or complexed via an optionally substitutedC₁-C₃₀ hydrocarbylene group or an optionally substituted C₃-C₃₀heterohydrocarbylene group.

Embodiment 15

The method of any of embodiments 12-14, wherein the NSAID or the residueof a NSAID is selected from the group consisting of acetylsalicylicacid, diflunisal, salsalate, choline magnesium trisalicylate, ibuprofen,dexibuprofen, naproxen, fenoprofen, ketoprofen, dexketoprofen,flurbiprofen, oxaprozin, loxoprofen, indomethacin, tolmetin, sulindac,etodolac, ketorolac, diclofenac, aceclofenac, nabumetone, piroxicam,meloxicam, tenoxicam, droxicam, lornoxicam, isoxicam, mefenamic acid,meclofenamic acid, flufenamic acid, tolfenamic acid, celecoxib,rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib,nimesulide, and licofelone, and a residue of any of the foregoingcompounds.

Embodiment 16

The method of any of embodiments 12-14, wherein the optionallysubstituted C₃-C₃₀ heterohydrocarbylene group is

where R¹ and R² are independently selected from hydrogen, C₁-C₄ alkyl,C₁-C₄ alkyl substituted with hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆cycloalkyl; or R¹ and R² together with the carbon to which they areattached form a C₃-C₈ cycloalkyl ring; and n is an integer selected from0 to 4, inclusive.

Embodiment 17

The method of embodiment 16, wherein R¹ and R² are methyl.

Embodiment 18

The method of embodiment 16, wherein the linking group is

Embodiment 19

The method of any of embodiments 12-14, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID is selected fromthe group consisting of naproxen, ketorolac, a residue of naproxen, anda residue of ketorolac.

Embodiment 20

The method of embodiment 12, wherein the conjugate is:

wherein Tc is ^(99m)Tc; or a pharmaceutically acceptable salt thereof.

Embodiment 21

The method of embodiment 12, wherein the conjugate is:

wherein Tc is ^(99m)Tc; or a pharmaceutically acceptable salt thereof.

Embodiment 22

The method of any of embodiments 12-14, 20, or 21, wherein the conjugateis administered as a composition additionally comprising apharmaceutically acceptable excipient.

Embodiment 23

A conjugate comprising: a non-steroidal anti-inflammatory drug (NSAID)or a residue of a NSAID bonded or complexed to an imaging moiety whichcomprises a radioactive agent, wherein the radioactive agent is selectedfrom the group consisting of a gamma-ray emitter and an X-ray emitter;or a pharmaceutically acceptable salt thereof.

Embodiment 24

The conjugate of embodiment 23, wherein said radioactive agent is^(99m)Tc, ⁵²Mn, ¹⁸⁶Re or ¹⁸⁸Re.

Embodiment 25

The conjugate of embodiment 23, of the form: (NSAID or NSAIDresidue)-(linker)-(chelating group)-M-(terminal ligand), where z is aninteger between 0 and 5 inclusive; and each “terminal ligand” isindependently selected from a ligand that bonds or complexes to M, butwhich does not have a NSAID or NSAID residue attached to it.

Embodiment 26

The conjugate of embodiment 23, wherein the NSAID or the residue of aNSAID and the imaging moiety are bonded or complexed via a linker,wherein the linker is selected from the group consisting of:

an optionally substituted C₁-C₄₀ hydrocarbylene group;

an optionally substituted C₂-C₄₀ heterohydrocarbylene group; and

a linker of the form -L_(E)-R⁴-L_(F)-,

-   -   where L_(E) is absent or is selected from the group consisting        of —NH— and —N(R⁸)—, and R⁸ is optionally substituted C₁-C₄        alkyl,    -   R⁴ is selected from the group consisting of optionally        substituted C₁-C₄₀ hydrocarbylene, optionally substituted C₂-C₄₀        heterohydrocarbylene, C₃-C₈ cycloalkyl, C alkyl-C₃-C₈        cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈        cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C alkyl-C₆-C₁₀ aryl, C₆-C₁₀        aryl-C₁-C₆ alkyl, and C₁-C₆ aryl-C₁-C₆ alkyl,    -   and L_(F) is absent or is a functional group selected from the        group consisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—,        —N(R⁹)—(C═O)—, —(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—,        —N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—, —(CH═CH)—, or a divalent        cycloalkyl or heterocyclic group, where R⁹ is selected from the        group consisting of H and optionally substituted C₁-C₄ alkyl.

Embodiment 27

The conjugate of embodiment 26, wherein the linker is selected from thegroup consisting of:

(—CH₂CH₂—O—)_(n) and (—CH₂CH(CH₃)—O—)_(n),where n is an integer from 1 to 12 inclusive.

Embodiment 28

The conjugate of embodiment 26, wherein the linker is selected from thegroup consisting of: —(NR_(a))—(CH₂)_(n)—(NR_(a))—,—(NH)—(CH₂)_(n)—(NH)—, —(NR_(a))—(CH₂CH₂)—(OCH₂CH₂)_(n)—(NR_(a))—,—(NH)—(CH₂CH₂)—(OCH₂CH₂)_(n)—(NH)—, where R_(a) is (C₁-C₄ alkyl) and nis an integer from 1 to 12 inclusive,

Embodiment 29

The conjugate of embodiment 23 or embodiment 24, wherein the imagingmoiety further comprises a group which bonds or complexes to theradioactive agent.

Embodiment 30

The conjugate of embodiment 29, wherein the group which bonds orcomplexes to the radioactive agent is a chelating group.

Embodiment 31

The conjugate of embodiment 30, wherein the chelating group is of theform:

wherein each J is independently selected from the group consisting of NHand S, R¹⁰¹, R¹⁰², and R¹⁰³ are independently selected from the groupconsisting of optionally substituted C₂-C₄ alkyl, andthe NSAID or NSAID residue is attached to theH-J-R¹⁰¹-J-R¹⁰²-J-R¹⁰³-J-H, either directly or through a linker, at anyJ, R¹⁰¹, R¹⁰², or R¹⁰³ atom where a hydrogen atom can be replaced with abond to the NSAID or NSAID residue, or to the linker.

Embodiment 32

The conjugate of embodiment 30, wherein the chelating group is selectedfrom the group consisting of:

where R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, and R¹⁷ are independentlyselected from the group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkylsubstituted with fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl;or, independently of the other substituents, (R¹⁰ and R¹¹) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, (R¹² and R¹³) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, or (R¹⁴ and R¹⁵) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, or (R¹⁶ and R¹⁷) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, with the proviso that only oneof (R¹⁰ and R¹¹), (R¹² and R¹³), (R¹⁴ and R¹⁵), and (R¹⁶ and R¹⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring; and wherein the bondmarked with an asterisk * indicates the attachment of the chelatinggroup to the remainder of the conjugate; and

R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³, R²⁴, and R²⁵ are independently selectedfrom the group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkylsubstituted with fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl;or, independently of the other substituents, (R¹⁸ and R¹⁹) together withthe carbon to which they are attached form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, (R²⁰ and R²¹) together with the carbon to whichthey are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,or (R²² and R²³) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁴ and R²⁵) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,with the proviso that only one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²²and R²³), or (R²⁴ and R²⁵) together with the carbon to which they areattached independently form a C₃-C₈ cycloalkyl ring or heterocycloalkylring; and wherein the bond marked with an asterisk * indicates theattachment of the chelating group to the remainder of the conjugate.

Embodiment 33

A conjugate of the formula:

[(NSAID or NSAID residue)-(linker)]_(x)-M-(terminal ligand)_(z)

where “NSAID or NSAID residue” refers to a NSAID or a residue of aNSAID; M is selected from the group consisting of ⁹⁹Tc, ⁵²Mn, and Re;x is an integer between 1 and 6 inclusive, z is an integer between 0 and5 inclusive, and the sum of x and z is less than or equal to 6; and“terminal ligand” is a ligand that coordinates to M, but which does nothave a NSAID attached to it; or a pharmaceutically acceptable saltthereof.

Embodiment 34

The conjugate of embodiment 33, wherein said conjugate is of theformula:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, where R⁸ is optionally substituted C₁-C₄ alkyl; R⁴ isselected from the group consisting of optionally substituted C₁-C₄₀hydrocarbylene, optionally substituted C₂-C₄₀ heterohydrocarbylene,C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀aryl-C₁-C₆ alkyl;L_(F) is absent or is a functional group selected from the groupconsisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O)N(R⁹)—, —(CH═CH)— or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl;where R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, and R¹⁷ are independentlyselected from the group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkylsubstituted with fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl;or, independently of the other substituents, (R¹⁰ and R¹¹) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, (R¹² and R¹³) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, or (R¹⁴ and R¹⁵) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, or (R¹⁶ and R¹⁷) together withthe carbon to which they are attached independently form a C₃-C₈cycloalkyl ring or heterocycloalkyl ring, with the proviso that only oneof (R¹⁰ and R¹¹), (R¹² and R¹³), (R¹⁴ and R¹⁵), and (R¹⁶ and R¹⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring; andM is selected from the group consisting of ^(99m)Tc and Re;or a pharmaceutically acceptable salt thereof.

Embodiment 35

The conjugate of embodiment 33, wherein said conjugate is of theformula:

(NSAID or NSAID residue)-(linker)-M-(terminal ligand)_(z)

and -(linker)-M-(terminal ligand)_(z) is selected from the groupconsisting of:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, and R⁸ is optionally substituted C₁-C₄ alkyl, with theproviso that if the L_(E) moiety of the group -L_(F)-R⁴-L_(E)-would beattached to a nitrogen atom, then L_(E) is absent;

where L_(G) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, and R⁸ is optionally substituted C₁-C₄ alkyl;

R⁴ is selected from the group consisting of optionally substitutedC₁-C₄₀ hydrocarbylene, optionally substituted C₂-C₄₀heterohydrocarbylene, C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl,C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl,C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆aryl-C₁-C₆ alkyl;

L_(F) is absent or is a functional group selected from the groupconsisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O) N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl; and R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³,R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, and R²⁹ are independently selected from thegroup consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted withfluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or, independentlyof the other substituents, (R¹⁸ and R¹⁹) together with the carbon towhich they are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkylring, (R²⁰ and R²¹) together with the carbon to which they are attachedform a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²² and R²³)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁴ and R²⁵)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁶ and R²⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁸ and R²⁹)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, with the proviso thatonly one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²² and R²³), or (R²⁴ andR²⁵), or (R²⁶ and R²⁷), or (R²⁸ and R²⁹) together with the carbon towhich they are attached independently form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring;

R³⁰, R³¹, R³², and R³³ are independently selected from hydrogen andC₁-C₄ alkyl, or one pair of (R³⁰ and R³¹), (R³¹ and R³²), or (R³² andR³³), together with the atoms to which they are attached, form asix-membered aryl ring or a five-to-six membered heteroaryl ring;R³⁴, R³⁵, R³⁶, and R³⁷ are independently selected from hydrogen andC₁-C₄ alkyl, or one pair of (R³⁴ and R³⁵), (R³⁵ and R³⁶), or (R³⁶ andR³⁷), together with the atoms to which they are attached, form asix-membered aryl ring or a five-to-six membered heteroaryl ring;R(_(ri)) is —CH₃ or —CH₂CH₃ and p is an integer between 0 and 4inclusive; and

X is Cl or Br;

or a pharmaceutically acceptable salt thereof.

Embodiment 36

The conjugate of embodiment 33, wherein said conjugate is of theformula:

(NSAID or NSAID residue)-(linker)-M-(terminal ligand)_(z)

and -(linker)-M-(terminal ligand)_(z) is selected from the groupconsisting of:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, and R⁸ is optionally substituted C₁-C₄ alkyl, with theproviso that if the L_(E) moiety of the group -L_(F)-R⁴-L_(E)-would beattached to a nitrogen atom, then L_(E) is absent;

R⁴ is selected from the group consisting of optionally substitutedC₁-C₄₀ hydrocarbylene, optionally substituted C₂-C₄₀heterohydrocarbylene, C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl,C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl,C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆aryl-C₁-C₆ alkyl;

L_(F) is absent or is a functional group selected from the groupconsisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O) N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl; and X is Cl or Br; or apharmaceutically acceptable salt thereof.

Embodiment 37

The conjugate of embodiment 33, wherein said conjugate is of theformula:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—,where R⁸ is optionally substituted C₁-C₄ alkyl,R⁴ is selected from the group consisting of optionally substitutedC₁-C₄₀ hydrocarbylene, optionally substituted C₂-C₄₀heterohydrocarbylene, C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl,C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl,C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆aryl-C₁-C₆ alkyl;R⁵ is selected from the group consisting of —OH, —NH₂, —NH(C₁-C₄ alkyl),and —N(C₁-C₄ alkyl)(C₁-C₄ alkyl);L_(F) is absent or is a functional group selected from the groupconsisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O)N(R⁹)—, —(CH═CH)— or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl;M is selected from the group consisting of ^(99m)Tc and Re; and R¹⁸,R¹⁹, R²⁰, R²¹, R²², R²³, R²⁴, and R²⁵ are independently selected fromthe group consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substitutedwith fluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or,independently of the other substituents, (R¹⁸ and R¹⁹) together with thecarbon to which they are attached form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, (R²⁰ and R²¹) together with the carbon to whichthey are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,or (R²² and R²³) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or(R²⁴ and R²⁵) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring,with the proviso that only one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²²and R²³), or (R²⁴ and R²⁵) together with the carbon to which they areattached independently form a C₃-C₈ cycloalkyl ring or heterocycloalkylring; or a pharmaceutically acceptable salt thereof.

Embodiment 38

The conjugate of any one of embodiments 23-26, wherein the linker isselected from the group consisting of:

where R¹, R², and R³ can be independently selected from the groupconsisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted withhydroxy, fluoro, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl;or R¹ and R² together with the carbon to which they are attached form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring; andn is an integer selected from 0 to 4, inclusive.

Embodiment 39

The conjugate of any one of embodiments 23-38, wherein the NSAID or theresidue of a NSAID is selected from the group consisting ofacetylsalicylic acid, diflunisal, salsalate, choline magnesiumtrisalicylate, ibuprofen, dexibuprofen, naproxen, fenoprofen,ketoprofen, dexketoprofen, flurbiprofen, oxaprozin, loxoprofen,indomethacin, tolmetin, sulindac, etodolac, ketorolac, diclofenac,aceclofenac, nabumetone, piroxicam, meloxicam, tenoxicam, droxicam,lornoxicam, isoxicam, mefenamic acid, meclofenamic acid, flufenamicacid, tolfenamic acid, celecoxib, rofecoxib, valdecoxib, parecoxib,lumiracoxib, etoricoxib, firocoxib, nimesulide, and licofelone, and aresidue of any of the foregoing compounds.

Embodiment 40

The conjugate of any one of embodiments 23-38, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID is selected fromthe group consisting of indomethacin, naproxen, ketorolac, celecoxib,rofecoxib, a residue of indomethacin, a residue of naproxen, a residueof ketorolac, a residue of celecoxib, and a residue of rofecoxib.

Embodiment 41

The conjugate of embodiment 40, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID is a residue of aNSAID selected from the group consisting of:

where the carbon atom or oxygen atom marked with an asterisk * indicatesan open valence at that atom at which the residue of the NSAID isattached to the remainder of the conjugate.

Embodiment 42

The conjugate of embodiment 40, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID is selected fromthe group consisting of indomethacin and a residue of indomethacin.

Embodiment 43

The conjugate of embodiment 40, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID is a residue of aNSAID selected from the group consisting of:

where the carbon atom or oxygen atom marked with an asterisk * indicatesan open valence at that atom at which the residue of the NSAID isattached to the remainder of the conjugate.

Embodiment 44

The conjugate of embodiment 23, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID bonded orcomplexed to an imaging moiety which comprises a radioactive agent isselected from the group consisting of:

wherein Tc is ^(99m)Tc;and pharmaceutically acceptable salts thereof.

Embodiment 45

The conjugate of embodiment 23, wherein the non-steroidalanti-inflammatory drug (NSAID) or a residue of a NSAID bonded orcomplexed to an imaging moiety which comprises a radioactive agent isselected from the group consisting of:

wherein Tc is ^(99m)Tc;and pharmaceutically acceptable salts thereof.

Embodiment 46

A pharmaceutical composition comprising one or more conjugates of anyone of embodiments 23-45, and a pharmaceutically acceptable excipient.

Embodiment 47

A method of imaging a site of pathology or suspected pathology in asubject, comprising:

a) administering one or more conjugates of any one of embodiments 23-45or the composition of embodiment 46 to the subject, wherein theradioactive agent of the conjugate comprises Re; and ^(99m)Tc, ⁵²Mn,¹⁸⁶Re or ¹⁸⁸Re; andb) generating an image of the subject or an image of a portion of thesubject.

Embodiment 48

The method of embodiment 47, wherein the pathology or suspectedpathology in the subject is a tumor or a suspected tumor.

Embodiment 49

The method of embodiment 47, wherein the subject is suffering from pain.

Embodiment 50

The method of embodiment 47, wherein the pathology or suspectedpathology in the subject is an infection or a suspected infection.

The disclosures of all publications, patents, patent applications andpublished patent applications referred to herein by an identifyingcitation are hereby incorporated herein by reference in their entirety.

The present invention has been described in terms of specificembodiments incorporating details to facilitate the understanding ofprinciples of construction and operation of the invention. Suchreference herein to specific embodiments and details thereof is notintended to limit the scope of the claims appended hereto. It will bereadily apparent to one skilled in the art that other variousmodifications can be made in the embodiments chosen for illustrationwithout departing from the spirit and scope of the invention. Therefore,the description and examples should not be construed as limiting thescope of the invention.

1. A conjugate of the formula:

where L_(E) is absent or is selected from the group consisting of —NH—,—N(R⁸)—, and —C(═O)—, and R⁸ is optionally substituted C₁-C₄ alkyl, R⁴is selected from the group consisting of optionally substituted C₁-C₁₀hydrocarbylene, optionally substituted C₂-C₁₀ heterohydrocarbylene,C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ aryl-C₁-C₆ alkyl,L_(F) is absent or is a functional group selected from the groupconsisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—,—(C═O)N(H)—, —N(H)—(C═O)—, —N(R⁹)—(C═O)—(CH₂)—, —(SO₂)N(R⁹)—,—N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—,—(CH═CH)—, or a divalent cycloalkyl or heterocyclic group, where R⁹ isselected from the group consisting of H and optionally substituted C₁-C₄alkyl; M is selected from the group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Reor ¹⁸⁸Re, where R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, and R¹⁷ areindependently selected from the group consisting of hydrogen, C₁-C₄alkyl, C₁-C₄ alkyl substituted with fluoro, hydroxy, —O—C₁-C₄ alkyl, orC₃-C₆ cycloalkyl; or, independently of the other substituents, (R¹⁰ andR¹¹) together with the carbon to which they are attached independentlyform a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, (R¹² and R¹³)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R¹⁴ and R¹⁵)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R¹⁶ and R¹⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, with the proviso thatonly one of (R¹⁰ and R¹¹) (R¹² and R¹³) (R¹⁴ and R¹⁵), and (R¹⁶ and R¹⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring; R₁₈, R₁₉, R₂₀, R₂₁, R₂₂,R₂₃, R₂₄, R₂₅, R₂₆, R₂₇, and R²⁹ are independently selected from thegroup consisting of hydrogen, C₁-C₄ alkyl, C₁-C₄ alkyl substituted withfluoro, hydroxy, —O—C₁-C₄ alkyl, or C₃-C₆ cycloalkyl; or, independentlyof the other substituents, (R¹⁸ and R¹⁹) together with the carbon towhich they are attached form a C₃-C₈ cycloalkyl ring or heterocycloalkylring, (R²⁰ and R²¹) together with the carbon to which they are attachedform a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²² and R²³)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁴ and R²⁵)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁶ and R²⁷)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, or (R²⁸ and R²⁹)together with the carbon to which they are attached independently form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, with the proviso thatonly one of (R¹⁸ and R¹⁹), (R²⁰ and R²¹), or (R²² and R²³), or (R²⁴ andR²⁵), or (R²⁶ and R²⁷), or (R²⁸ and R²⁹) together with the carbon towhich they are attached independently form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring; or, independently of the other substituents, (R²²and R²³) together form an oxo group; where Q is selected from the groupconsisting of OH, NH₂, NH(C₁-C₄ alkyl), N(C₁-C₄ alkyl)(C₁-C₄ alkyl),OCH₃, and OCH₂CH₃; or a pharmaceutically acceptable salt thereof,wherein the NSAID or NSAID residue or NSAID derivative is celecoxib, aresidue of celecoxib, or a derivative of celecoxib. 2-5. (canceled)
 6. Aconjugate comprising: a non-steroidal anti-inflammatory drug (NSAID), aresidue of a NSAID, or a derivative of a NSAID bonded or complexed to animaging moiety which comprises a radioactive agent, wherein theradioactive agent is selected from the group consisting of a gamma-rayemitter, an X-ray emitter, and a beta emitter; or a pharmaceuticallyacceptable salt thereof, wherein the non-steroidal anti-inflammatorydrug (NSAID), residue of a NSAID, or derivative of a NSAID is celecoxib,a residue of celecoxib, or a derivative of celecoxib.
 7. The conjugateof claim 6, wherein said radioactive agent is ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re or¹⁸⁸Re.
 8. The conjugate of claim 7, wherein the imaging moiety furthercomprises a chelating group which bonds or complexes to the radioactiveagent; and where the imaging moiety is optionally bonded to thenon-steroidal anti-inflammatory drug (NSAID), residue of a NSAID, orderivative of a NSAID via a linker.
 9. The conjugate of claim 8, whereinthe imaging moiety comprising a chelating group bonded or complexed tothe radioactive agent is of the form:

wherein M is selected from the group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Reor ¹⁸⁸Re, wherein each J is independently selected from the groupconsisting of NH and S, and R¹⁰¹, R¹⁰², and R¹⁰³ are independentlyselected from the group consisting of optionally substituted C₂-C₄alkyl, and the NSAID, NSAID residue, or NSAID derivative is attached tothe chelating group, either through a linker or directly, at a) any J,R¹⁰¹, R¹⁰², or R¹⁰³ atom where a hydrogen atom can be replaced with abond to the linker (if present) or to the NSAID, NSAID residue, or NSAIDderivative if no linker is present; or at b) the nitrogen atom in the—R¹⁰¹—N—R¹⁰²— portion, forming a bond between that nitrogen and thelinker (if present) or to the NSAID, NSAID residue, or NSAID derivativeif no linker is present; or at c) the nitrogen atom in the —R¹⁰²—N—R¹⁰³—portion, forming a bond between that nitrogen and the linker (ifpresent) or to the NSAID, NSAID residue, or NSAID derivative if nolinker is present; or wherein the imaging moiety comprising a chelatinggroup bonded or complexed to the radioactive agent is of the form:

wherein M is selected from the group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Reor ¹⁸⁸Re, wherein R_((ri)) is —CH₃ or —CH₂CH₃ and p is an integerbetween 0 and 4 inclusive, and the NSAID, NSAID residue, or NSAIDderivative is attached to the chelating group either through a linker ordirectly if no linker is present, at any location on the cyclopentanering which does not have a (R_(ri)) group.
 10. The conjugate of claim 8,wherein the NSAID, residue of an NSAID, or derivative of a NSAID and theimaging moiety are bonded or complexed via a linker, wherein the linkeris selected from the group consisting of: an optionally substitutedC₁-C₁₀ hydrocarbylene group; an optionally substituted C₂-C₁₀heterohydrocarbylene group; and a linker of the form -L_(E)-R⁴-L_(F)-;where L_(E) is absent or is selected from the group consisting of —NH—,—N(R⁸)—, and —C(═O)—, and R⁸ is optionally substituted C₁-C₄ alkyl, R⁴is selected from the group consisting of optionally substituted C₁-C₁₀hydrocarbylene, optionally substituted C₂-C₁₀ heterohydrocarbylene,C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀aryl-C₁-C₆ alkyl, and L_(F) is absent or is a functional group selectedfrom the group consisting of —(C═O)—, —O—, —N(R⁹)—, —(C═O)N(R⁹)—,—N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—, —N(R⁹)—(C═O)—(CH₂)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O)N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl.
 11. The conjugate of claim 8,wherein the linker is selected from the group consisting of:—(NH)—(CH₂)_(n), —(NR_(a))—(CH₂)_(n)—, —(NH)—(CH₂)_(n)—(NH)—,—(NR_(a))—(CH₂)_(n)—(NR_(a))—, —(NH)—(CH₂CH₂)—(OCH₂CH₂)_(m)—(NH)—,—(NR_(a))—(CH₂CH₂)—(OCH₂CH₂)_(m)—(NR_(a))—,—(NH)—CH₂CH₂)—((NH)CH₂CH₂)_(m)—(NH)—,—(NR_(a))—(CH₂CH₂)—((NH)CH₂CH₂)_(m)—(NR_(a))—(—CH₂CH₂—O—)_(n),(—CH₂CH(CH₃)—O—)_(q), where R_(a) is (C₁-C₄ alkyl), n is an integer from1 to 10 inclusive, m is an integer from 1 to 4 inclusive, and q is aninteger from 1 to 3 inclusive, —NH—(CH₂)₂—, —NH—(CH₂)₃—, —NH—(CH₂)₄—,—NH—(CH₂)₅—, —NH—(CH₂)₆—, NH—(CH₂)₇—, —NH—(CH₂)₈—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—, —NH—(CH₂)₂—NH—, —NH—(CH₂)₃—NH—,—NH—(CH₂)₄—NH—, —NH—(CH₂)₅—NH—, —NH—(CH₂)₆—NH—, —NH—(CH₂)₇—NH—,—NH—(CH₂)₈—NH—, —NH—(CH₂)₃—C(CH₃)₂—NH—, —NH—(CH₂)₄—C(CH₃)₂—NH—,—NH—(CH₂)₅—C(CH₃)₂—NH—, —NH—(CH₂)₆—C(CH₃)₂—NH—, —NH—(CH₂)₇—C(CH₃)₂—NH—,—NH—(CH₂)₂—C(CH₃)₂—(CH₂)₂—NH—, —NH—(CH₂)₂—C(CH₃)₂—(CH₂)₂—,—NH—C(CH₃)₂—(CH₂)₃—, —NH—C(CH₃)₂—(CH₂)₄—, —NH—C(CH₃)₂—(CH₂)₅—,—NH—C(CH₃)₂—(CH₂)₆—, —NH—C(CH₃)₂—(CH₂)₇—, —NH—CH(CH₃)—(CH₂)₃—,—NH—CH(CH₃)—(CH₂)₄—, —NH—CH(CH₃)—(CH₂)₅—, —NH—CH(CH₃)—(CH₂)₆—,—NH—CH(CH₃)—(CH₂)₇—, —NH—CH(CF₃)—(CH₂)₃—, —NH—CH(CF₃)—(CH₂)₄—,—NH—CH(CF₃)—(CH₂)₅—, —NH—CH(CF₃)—(CH₂)₆—, —NH—CH(CF₃)—(CH₂)₇—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—, —NH—(CH₂)₂—NH—(C═O)—,—NH—(CH₂)₃—NH—(C═O)—, —NH—(CH₂)₄—NH—(C═O)—, —NH—(CH₂)₅—NH—(C═O)—,—NH—(CH₂)₆—NH—(C═O)—, —NH—(CH₂)₇—NH—(C═O)—, —NH—(CH₂)₈—NH—(C═O)—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—(C═O)—, —NH—(CH₂)₂—NH—(C═O)—(CH₂)—,—NH—(CH₂)₃—NH—(C═O)—(CH₂)—, —NH—(CH₂)₄—NH—(C═O)—(CH₂)—,NH—(CH₂)₅—NH—(C═O)—(CH₂)—, —NH—(CH₂)₆—NH—(C═O)—(CH₂)—,—NH—(CH₂)₇—NH—(C═O)—(CH₂)—, —NH—(CH₂)₈—NH—(C═O)—(CH₂)—,—NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—(C═O)—(CH₂)—, —C(═O)—(CH₂)₂—,—C(═O)—(CH₂)₃—, —C(═O)—(CH₂)₄—, —C(═O)—(CH₂)₅—, —C(═O)—(CH₂)₆—,—C(═O)—(CH₂)₇—, —C(═O)—(CH₂)₈—, —C(═O)—(CH₂)₂—C(H)═C(H)—(CH₂)₂—,—C(═O)—(CH₂)₂—NH—, —C(═O)—(CH₂)₃—NH—, —C(═O)—(CH₂)₄—NH—,—C(═O)—(CH₂)₅—NH—, —C(═O)—(CH₂)₆—NH—, —C(═O)—(CH₂)₇—NH—,—C(═O)—(CH₂)₈—NH—, —C(═O)—(CH₂)₂—C(H)═C(H)—(CH₂)₂—NH—,—NH—(CH₂)₂—O—(CH₂)₂—NH—, —NH—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—NH—,—NH—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—NH—, —NH—(CH₂)₂—NH—(CH₂)₂—NH—,—NH—(CH₂)₂—NH—(CH₂)₂—NH—(CH₂)₂—NH—,—NH—(CH₂)₂—NH—(CH₂)₂—NH—(CH₂)₂—NH—(CH₂)₂—NH—, —NH—(CH₂)₂—N(CH₃)—,—NH—(CH₂)₃—N(CH₃)—, —NH—(CH₂)₄—N(CH₃)—, —NH—(CH₂)₅—N(CH₃)—,—NH—(CH₂)₆—N(CH₃)—, —NH—(CH₂)₇—N(CH₃)—, —NH—(CH₂)₈—N(CH₃)—,—NH—(CH₂)₃—NH—(CH₂)₃—, —NH—(CH₂)₂—C(H)═C(H)—(CH₂)₂—,—NH—CH₂—CF₂—(CH₂)₄—, —NH—CH₂—CF₂—(CH₂)₅— C(═O)—CF₂—(CH₂)₄—,C(═O)—CF₂—(CH₂)₅—,


12. The conjugate of claim 1, wherein said conjugate is of the formula:(NSAID, NSAID residue, or NSAIDderivative)-(linker)-(chelator)-M-(terminal ligand)_(z1) or (NSAID,NSAID residue, or NSAID derivative)-(linker)-M-(terminal ligand)_(z2)where z1 is an integer between 0 and 4 inclusive; z2 is an integerbetween 0 and 5 inclusive; and -(linker)-(chelator)-M-(terminalligand)_(z1) or -(linker)-M-(terminal ligand)_(z2) is selected from thegroup consisting of:

where L_(E) is absent or is selected from the group consisting of —NH—,—N(R⁸)—, and —(C═O)—, and R⁸ is optionally substituted C₁-C₄ alkyl, withthe proviso that if the L_(E) moiety of the group -L_(F)-R⁴-L_(E)-wouldbe attached to a nitrogen atom, then L_(E) is absent; where L_(G) isabsent or is selected from the group consisting of —NH—, —N(R⁸)—, and—(C═O)—, and R⁸ is optionally substituted C₁-C₄ alkyl; R⁴ is selectedfrom the group consisting of optionally substituted C₁-C₁₀hydrocarbylene, optionally substituted C₂-C₁₀ heterohydrocarbylene,C₃-C₈ cycloalkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆alkyl, C₁-C₆ alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆alkyl-C₆-C₁₀ aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ alkyl-C₆-C₁₀aryl-C₁-C₆ alkyl; R⁵ is selected from the group consisting of —OH, —NH₂,—NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —OCH₃, and —OCH₂CH₃;R_(N) is H or (C₁-C₄ alkyl); L_(F) is absent or is a functional groupselected from the group consisting of —(C═O)—, —O—, —N(R⁹)—,—(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—, —(SO₂)N(R⁹)—,—N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—, —O—(C═O)N(R⁹)—,—(CH═CH)—, or a divalent cycloalkyl or heterocyclic group, where R⁹ isselected from the group consisting of H and optionally substituted C₁-C₄alkyl; and R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, andR²⁹ are independently selected from the group consisting of hydrogen,C₁-C₄ alkyl, C₁-C₄ alkyl substituted with fluoro, hydroxy, —O—C₁-C₄alkyl, or C₃-C₆ cycloalkyl; or, independently of the other substituents,(R¹⁸ and R¹⁹) together with the carbon to which they are attached form aC₃-C₈ cycloalkyl ring or heterocycloalkyl ring, (R²⁰ and R²¹) togetherwith the carbon to which they are attached form a C₃-C₈ cycloalkyl ringor heterocycloalkyl ring, or (R²² and R²³) together with the carbon towhich they are attached independently form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, or (R²⁴ and R²⁵) together with the carbon towhich they are attached independently form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, or (R²⁶ and R²⁷) together with the carbon towhich they are attached independently form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, or (R²⁸ and R²⁹) together with the carbon towhich they are attached independently form a C₃-C₈ cycloalkyl ring orheterocycloalkyl ring, with the proviso that only one of (R¹⁸ and R¹⁹),(R²⁰ and R²¹), or (R²² and R²³), or (R²⁴ and R²⁵), or (R²⁶ and R²⁷), or(R²⁸ and R²⁹) together with the carbon to which they are attachedindependently form a C₃-C₈ cycloalkyl ring or heterocycloalkyl ring;R³⁰, R³¹, R³², and R³³ are independently selected from hydrogen andC₁-C₄ alkyl, or one pair of (R³⁰ and R³¹), (R³¹ and R³²), or (R³² andR³³), together with the atoms to which they are attached, form asix-membered aryl ring or a five-to-six membered heteroaryl ring; R³⁴,R³⁵, R³⁶, and R³⁷ are independently selected from hydrogen and C₁-C₄alkyl, or one pair of (R³⁴ and R³⁵), (R³⁵ and R³⁶), or (R³⁶ and R³⁷),together with the atoms to which they are attached, form a six-memberedaryl ring or a five-to-six membered heteroaryl ring; M is selected fromthe group consisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re and ¹⁸⁸Re; R(_(ri)) is—CH₃ or —CH₂CH₃ and p is an integer between 0 and 4 inclusive; and X isCl or Br; or a pharmaceutically acceptable salt thereof.
 13. Theconjugate of claim 12, wherein -(linker)-(chelator)-M-(terminalligand)_(z1) or -(linker)-M-(terminal ligand)_(z2) is selected from thegroup consisting of:

where L_(E) is absent or is selected from the group consisting of —NH—and —N(R⁸)—, and R⁸ is optionally substituted C₁-C₄ alkyl, with theproviso that if the L_(E) moiety of the group -L_(F)-R⁴-L_(E)-would beattached to a nitrogen atom, then L_(E) is absent; R⁴ is selected fromthe group consisting of optionally substituted C₁-C₁₀ hydrocarbylene,optionally substituted C₂-C₁₀ heterohydrocarbylene, C₃-C₈ cycloalkyl,C₁-C₆ alkyl-C₃-C₈ cycloalkyl, C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₁-C₆alkyl-C₃-C₈ cycloalkyl-C₁-C₆ alkyl, C₆-C₁₀ aryl, C₁-C₆ alkyl-C₆-C₁₀aryl, C₆-C₁₀ aryl-C₁-C₆ alkyl, and C₁-C₆ aryl-C₁-C₆ alkyl; R⁵ isselected from the group consisting of —OH, —NH₂, —NH(C₁-C₄ alkyl),—N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —OCH₃, and —OCH₂CH₃; L_(F) is absent or isa functional group selected from the group consisting of —(C═O)—, —O—,—N(R⁹)—, —(C═O)N(R⁹)—, —N(R⁹)—(C═O)—, —(C═O)N(H)—, —N(H)—(C═O)—,—(SO₂)N(R⁹)—, —N(R⁹)—(SO₂)—, —N(R⁹)(C═O)N(R⁹)—, —N(R⁹)—(C═O)—O—,—O—(C═O)N(R⁹)—, —(CH═CH)—, or a divalent cycloalkyl or heterocyclicgroup, where R⁹ is selected from the group consisting of H andoptionally substituted C₁-C₄ alkyl; M is selected from the groupconsisting of ^(99m)Tc, ⁵²Mn, ¹⁸⁶Re and ¹⁸⁸Re; and X is Cl or Br; or apharmaceutically acceptable salt thereof.
 14. The conjugate of claim 1,wherein the non-steroidal anti-inflammatory drug (NSAID), residue of aNSAID, or derivative of a NSAID is selected from the group consistingof:

where the atom marked with an asterisk * indicates an open valence atthat atom at which the NSAID, residue of the NSAID or derivative of theNSAID is attached to the remainder of the conjugate. 15-16. (canceled)17. The conjugate of claim 1, which is selected from the groupconsisting of:

wherein Tc is ^(99m)Tc; and pharmaceutically acceptable salts thereof.18. (canceled)
 19. A pharmaceutical composition comprising one or moreconjugates of claim 1, and a pharmaceutically acceptable excipient. 20.A method of imaging a site of pathology or suspected pathology in asubject, comprising: a) administering one or more conjugates of claim 1to the subject, wherein the radioactive agent of the conjugate comprises^(99m)Tc, ⁵²Mn, ¹⁸⁶Re or ¹⁸⁸Re; and b) generating an image of thesubject or an image of a portion of the subject.
 21. The method of claim20, wherein the pathology or suspected pathology in the subject is atumor or a suspected tumor.
 22. The method of claim 20, wherein thesubject is suffering from pain.
 23. The method of claim 20, wherein thepathology or suspected pathology in the subject is an infection or asuspected infection.
 24. The conjugate of claim 4, wherein saidconjugate has an IC₅₀ for cyclooxygenase inhibition of less than about0.5 micromolar.
 25. The conjugate of claim 24, wherein thecyclooxygenase is COX-2.